Classification of CMD according to the involvement of pathogenic mechanisms and the clinical setting.
\r\n\t
",isbn:"978-1-80356-357-2",printIsbn:"978-1-80356-356-5",pdfIsbn:"978-1-80356-358-9",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,hash:"3aba1eb3600a8c9ff880c628f70b3298",bookSignature:"Ph.D. Delfín Ortega-Sánchez",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11481.jpg",keywords:"Integrated Curriculum, Transdisciplinarity, Integrated Active Learning, Educational Programs, Contemporary Social Problems, Critical Thinking, Creative Thinking, Social Thinking, Agenda 2030, Sustainable Development Goals, Educational Paradigm, Social Reality",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"February 18th 2022",dateEndSecondStepPublish:"March 18th 2022",dateEndThirdStepPublish:"May 17th 2022",dateEndFourthStepPublish:"August 5th 2022",dateEndFifthStepPublish:"October 4th 2022",remainingDaysToSecondStep:"2 months",secondStepPassed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Internationally recognized researcher in the field of historical and social science education. Author of more than 100 publications, awarded three Doctorate degrees and the National End of Degree Award, granted by the Ministry of Education to the best academic records of Bachelor's degrees in Spain. Dr. Ortega-Sánchez has been Vice-Rector for Social Responsibility, Culture, and Sports at the University of Burgos since 2021.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"302925",title:"Ph.D.",name:"Delfín",middleName:null,surname:"Ortega-Sánchez",slug:"delfin-ortega-sanchez",fullName:"Delfín Ortega-Sánchez",profilePictureURL:"https://mts.intechopen.com/storage/users/302925/images/system/302925.jpg",biography:"I hold a PhD in Didactics of Social Sciences from the Autonomous University of Barcelona, a PhD in Educational Sciences from the University of Burgos, and a PhD in History from the University of Extremadura. My research interests focus on the construction of identities in the History and Geography teaching, gender mainstreaming in initial education and training for teachers, the didactic treatment of relevant social problems and controversial issues in the teaching of the social and human sciences, and the application of educational technology in the specific field of social sciences. I am currently a Social Sciences teacher and researcher at University of Burgos (Spain).",institutionString:"University of Burgos",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"University of Burgos",institutionURL:null,country:{name:"Spain"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"23",title:"Social Sciences",slug:"social-sciences"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"429339",firstName:"Jelena",lastName:"Vrdoljak",middleName:null,title:"Ms.",imageUrl:"https://mts.intechopen.com/storage/users/429339/images/20012_n.jpg",email:"jelena.v@intechopen.com",biography:"As an Author Service Manager, my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. 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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"72",title:"Ionic Liquids",subtitle:"Theory, Properties, New Approaches",isOpenForSubmission:!1,hash:"d94ffa3cfa10505e3b1d676d46fcd3f5",slug:"ionic-liquids-theory-properties-new-approaches",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/72.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"314",title:"Regenerative Medicine and Tissue Engineering",subtitle:"Cells and Biomaterials",isOpenForSubmission:!1,hash:"bb67e80e480c86bb8315458012d65686",slug:"regenerative-medicine-and-tissue-engineering-cells-and-biomaterials",bookSignature:"Daniel Eberli",coverURL:"https://cdn.intechopen.com/books/images_new/314.jpg",editedByType:"Edited by",editors:[{id:"6495",title:"Dr.",name:"Daniel",surname:"Eberli",slug:"daniel-eberli",fullName:"Daniel Eberli"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"57",title:"Physics and Applications of Graphene",subtitle:"Experiments",isOpenForSubmission:!1,hash:"0e6622a71cf4f02f45bfdd5691e1189a",slug:"physics-and-applications-of-graphene-experiments",bookSignature:"Sergey Mikhailov",coverURL:"https://cdn.intechopen.com/books/images_new/57.jpg",editedByType:"Edited by",editors:[{id:"16042",title:"Dr.",name:"Sergey",surname:"Mikhailov",slug:"sergey-mikhailov",fullName:"Sergey Mikhailov"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1373",title:"Ionic Liquids",subtitle:"Applications and Perspectives",isOpenForSubmission:!1,hash:"5e9ae5ae9167cde4b344e499a792c41c",slug:"ionic-liquids-applications-and-perspectives",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/1373.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"51938",title:"The Morphology, Physiology and Pathophysiology of Coronary Microcirculation",doi:"10.5772/64537",slug:"the-morphology-physiology-and-pathophysiology-of-coronary-microcirculation",body:'\nThe heart is one of the most demanding organs of the human body as it presents high demands for nutrients and oxygen. These demands are physiologically met through an extensive and unique vascular network, which is usually known as
Historically, the large epicardial arteries were considered the coronary circulation. Nowadays, the scientific reports suggest the coronary circulation is characterized by an extreme complexity in terms of morphology but also physiology. Moreover, the theory that the coronary circulation involves the larger epicardial arteries is no longer acceptable given the extensive vascular network present in the myocardium.
\nCoronary artery disease (CAD) is usually associated with larger epicardial coronary arteries. However, previous studies have shown an important link between microcirculatory dysfunction and cardiovascular disease. In fact, pathological changes in smaller vessels have been detected prior to clinical manifestations of cardiovascular disease [1]. Moreover, microcirculatory dysfunction may even be a risk indicator for metabolic syndrome and associated cardiovascular disease [1, 2].
\nThis review aimed to explore the (a) morphology, with particular interest on the anatomical and histological aspects; (b) physiology, providing an insight on the several endothelium-dependent and endothelium-independent regulatory mechanisms; and (c) pathophysiology of the cardiac microcirculation, with a special focus on the changes in the regulatory mechanisms, on the atherogenesis and on the correlation to the systemic cardiovascular disease.
\nBased on the morphology and function, the coronary circulation involves several types of vessels as follows (from the larger arteries to the largest veins): epicardial arteries or coronary arteries, small arteries or intramural arteries, arterioles, capillaries, venules and epicardial veins. These vessels may be grouped according to their size into (a)
The
The anatomy of these vessels varies along their length: the proximal and middle portions tend to present similar characteristics to the larger arteries, although with a thick tunica media [several layers of vascular smooth muscle cells (VSMCs)], while the distal portions, also termed terminal or precapillary arterioles, may present a thinner tunica media (one to two layers of VSMCs) or not even present any VSMC layer, which is replaced by small unique cells that will be explored in the following subsection, the
Vascular network and capillary neurovascular unit. As presented in the figure, the larger arteries (with a well-defined smooth muscle layer that may vary in size) present morphological differences to the capillaries, which do not present smooth muscle layer, being substituted by pericytes. The capillary neurovascular unit then includes the endothelium, basal lamina and pericytes, which are surrounded by neuron terminals. Moreover, the vascular network also includes other cell types, such as fibroblasts, collateral blood vessels, among others. Adapted from Zhang et al. [
The connection between the arterial and the venous systems is fundamentally achieved by a capillary network placed amid the arterioles and the venules (Figure 1).
\nThe
These vessels present structural differences to other vessels as the wall is essentially composed of two layers: an inner layer, the
According to their morphology, capillaries may be classified into three main categories: (a) continuous capillaries, (b) fenestrated capillaries and (c) discontinuous capillaries [6]. In the coronary microcirculation, the
Embedded in the basal membrane of capillaries, between the endothelium and the parenchyma, small contractile cells called
Pericytes may vary morphologically and physiologically depending on the vascular bed and on the position in the vascular bed itself [13]. Nevertheless, they generally extend processes along and around capillaries [12, 13]. In the central nervous system and kidneys, pericytes play an important role in angiogenesis, regulation of the endothelium, among other functions [12, 13]. These cells seem to be particularly relevant in the central nervous system where the regional blood flow regulation is of crucial importance [13]. These pericytes may also present contractile properties [12, 13]. Several proteins have been suggested to confer contractility to pericytes, such as α-smooth muscle actin and tropomyosin [13]. However, previous studies suggest that the contractile mechanisms differ from the VSMCs [13].
\nAlthough the role of pericytes in coronary physiology is not yet fully understood, the high number of these cells in cardiac capillaries and the similar characteristics to the central nervous system pericytes indicates these cells may play an important role in the regulation of the vessel diameter as well as permeability [12].
\nIn the capillary network, other structures may be found such as
After the exchange of nutrients and oxygen at the capillary level, the deoxygenated blood, containing metabolic products, proceeds to the
The venules usually present a diameter ranging from 10 to 50 μm and similar anatomical characteristics to the arterioles [8]. The proximal venules, that is postcapillary venules, usually exhibit only two layers: an inner layer, the
The distal venules are morphologically different relatively to the postcapillary venules, as they may present a thin tunica media (one to two layers of VSMCs) and a thin tunica adventitia on the outer side of the vessel [6]. The absence of pericytes is a key characteristic of these distal venules [6]. These venules initially course parallel to the muscle fibres, accompanying the arterioles and capillaries, then changing their position and configuration to meet the larger coronary veins [5].
\nIn healthy conditions, the myocardial blood supply is fundamentally provided through the normal coronary circulation. However, in the presence of cardiac disease, such as chronic cardiac disease or regional ischemic injuries, the myocardial perfusion may be compromised [4, 5]. Compensatory circulatory communications named
The direct communication between the heart chamber and the coronary circulation is generally referred to
The physiologic behaviour of the coronary circulation is inherently linked to a balance between the blood supply and the metabolic demand of the heart [19]. Furthermore, the physiological responses in the microcirculation seem to depend on the vessel size and type and appear to vary within the microcirculation itself and from those in the macrocirculation [19–21]. Physiologically, the coronary microcirculation is able to respond to a wide range of stimuli, such as growth and physical exercise, through adaptive processes, essential to the maintenance of its physiology [19]. In fact, vessels present a high adaptation ability and may undergo both acute and chronic adjustments. The acute adjustments involve changes in the vascular smooth muscle tone, while the chronic adjustments involve wall structure changes [19].
\nThe
The
The coronary blood flow is intrinsically linked with metabolic demands of the myocardium, namely of oxygen. At rest, the myocardial oxygen extraction averages 60–70%, which leads to the coronary venous pO2 of about 20 mmHg [28]. During physical exercise, several mechanisms of adaptation are triggered in the myocardium, pO2 seems to be kept constant, which highlights the role of several pathways, namely the myocardial aerobic metabolism [28]. This energy production is generally dependent on mitochondrial oxidative phosphorylation pathways [19]. Among several metabolites produced in these intracellular pathways, carbon dioxide (CO2) and reactive oxygen species (ROS) seem to play an important role in physiological conditions [19, 28].
\nAs previously mentioned,
After the conversion of oxaloacetate into citrate, the citric acid cycle involves several reactions in chain. Some of them also involve the production of CO2, such as the production of α-ketoglutarate (reaction 2) and succinyl-CoA (reaction 3).
The increased production of CO2 may also induce a decrease in pH due to the increase in proton concentration, as presented in the following reaction:
This change in pH seems to promote the coronary vasodilation [28–30].
\nAs can be seen in Figure 2, the metabolic production of
The vasodilator properties of H2O2 have long been studied, but the precise underlying mechanisms are not yet fully established [28]. Previous studies suggested H2O2 behaves as an endothelium-derived hyperpolarizing factor (EDHF) [34, 35], as described in the following subsection. However, other previous studies suggested the mechanism may involve the stimulation of the nitric oxide (NO) production or be mediated by the guanylyl cyclase in human coronary arterioles [36]. These pathways will be further discussed in the following subsections.
\nOther studies have suggested additional mechanisms involved in the metabolic regulation exerted by H2O2 on the coronary blood flow. The involvement of oxidation of thiol groups as a pathway of coronary metabolic dilation in isolated coronary arterioles has been previously proposed [37]. The thiol groups are involved in many pathophysiological mechanisms and play a key role in the biological protection against oxidative injuries [38, 39]. This oxidation process promotes modifications in the protein conformation and includes the conversion of protein-bound thiols (-SH) into sulfenic (SO−, reaction 5), sulphinic (SOO−) and sulphonic (SOOO−) acids as well as disulphide bridges (S-S, reaction 6) [37, 39].
\nFeedforward reactive oxygen species-dependent metabolic regulation of coronary blood flow. The increased metabolic demands of the myocardium trigger an increase in mitochondrial metabolism and flux, through the electron transport chain (ETC), increasing the production of O2⋅− and subsequently of H2O2 by manganese SOD (MnSOD). H2O2 then diffuses to the VSMC and activates voltage-dependent K+ (KV) channels promoting the hyperpolarization of the VSMCs and thus the vasodilation in the coronary microcirculation. Adapted from Muller-Delp [
Furthermore, these modifications in the redox state of the cell may also affect the hyperpolarization mediated by thiol-dependent voltage-dependent K+ (KV) channels, which will be further explored in the following subsection [40].
\nOther metabolic vasodilators may also be involved, such as adenosine (which concentration is dependent on the metabolism) and potassium ions, which will be explored further below.
\nThe
Multitude of pathways involved in the endothelium-dependent contraction. Abbreviations: 5-HT, 5-hydrotryptamine; ACE, angiotensin-converting enzyme; ACh, acetylcholine; ADP, adenosine diphosphate; AT-I, angiotensin-I; AT1, angiotensin receptor; AT-II, angiotensin-II; ATG, angiotensinogen; BDK, bradykinin; COX, cyclooxygenases; ECE, endothelin-converting enzyme; ET-1, endothelin-1; ETA, endothelin receptor A; ETB, endothelin receptor B; NO, nitric oxide; eNOS, endothelial nitric oxide synthase; NOX, NADPH oxidase; PGs, prostaglandins; PLA2, phospholipase A2; ROS, reactive oxygen species; TGFβ1, transforming growth factor; Thr, thrombin; TP, thromboxane-prostanoid receptor; TXA2, thromboxane A2; VSMC, vascular smooth muscle cell; XO, xanthine oxidase. Adapted from Virdis et al. [
Pathways involved in the endothelium-dependent and endothelium-independent relaxation of the VSMC. Stimulation of the endothelial cells by acetylcholine (ACh) or other agents (e.g. bradykinin and shear stress) results in the formation and release of an EDRF identified as nitric oxide (NO). Substances such as adenosine, nitroprusside (NP), H+, CO2 and K+ can be produced in the parenchymal tissue and elicit vasodilation by direct action on vascular smooth muscle. Adapted from Koeppen et al. [
Although NO is considered the major pathway of endothelium-mediated vasodilation in the systemic circulation, multiple pathways may be involved in this physiological response, such as the prostaglandins-induced vasodilation (Figure 4). The
Several vasoactive substances have been included in the
Hyperpolarization of the VSMC. Abbreviations: ACh, acetylcholine; BK, bradykinin; BKCa, large conductance Ca2+-activated K+ channels; CaV, voltage-activated Ca2+ channels; Cx, connexin; EC, endothelial cell; eNOS, endothelial nitric oxide synthase; IKCa, intermediate conductance Ca2+-activated K+ channels; KIR, inwardly rectifying K+ channels; NO, nitric oxide; PE, phenylephrine; RyR, ryanodine receptor; SKCa, small conductance Ca2+-activated K+ channels; SP, substance P, TRPC1, transient receptor potential canonical channel 1; TRPV4, transient receptor potential vanilloid channel 4; VSMC, vascular smooth muscle cell. Adapted from Félétou et al. [
The hyperpolarization of the VSMC may involve several ionic channels, such as the voltage-activated Ca2+ (CaV) channels, which regulate the intracellular Ca2+ concentration, the KV channels and the Ca2+-activated K+ (KCa) channels [35, 40]. The KCa channels may be subdivided into small (SKCa or KCa 2.3 isoform), intermediate (IKCa or KCa 3.1 isoform) and large (BKCa) conductance Ca2+-activated K+ channels, which are located in specific cellular and subcellular sites [35]. The hyperpolarization of the VSMCs may be triggered directly, through receptors on the VSMC membrane, or indirectly, through the hyperpolarization of the endothelial cells [35].
\nAs can be seen in Figure 5, the
Moreover, the VSMCs may be
Besides the hyperpolarization of the VSMCs, the EDHFs, particularly H2O2, may also promote vasodilation through other mechanisms, namely by stimulating the production of prostaglandin E2 in the endothelial cell, thus promoting the endothelium-dependent vasodilation [62].
\nThe relative importance of each pathway is still unestablished, but it has been proposed to depend for example on the activation state of the VSMCs, the density of MEJs and the expression of KIR and Na+/K+-ATPase [57].
\nAs previously mentioned, in addition to the stimulation of receptors on the endothelial cell membrane, other factors may modulate the endothelial function, namely the forces exerted by the blood flow on the vessel wall. There are two major forces: (a) one perpendicular to the wall and (b) another parallel to the wall, known as
Flow-mediated dilation in the human coronary arterioles. Abbreviations: AA, arachidonic acid; BKCa, large conductance Ca2+-activated K+ channels; cGMP, cyclic guanosine monophosphate; CuZnSOD, copper-zinc superoxide dismutase; CYP, cytochrome P450; CYS 42, cysteine residue; EETs, epoxyeicosatrienoic acids; GC, guanylyl cyclase; GTP; guanosine triphosphate; H2O2, hydrogen peroxide; KCa, Ca2+-activated K+ channels; MnSOD, manganese superoxide dismutase; NADPH, nicotinamide adenine dinucleotide phosphate; NOX, NADPH oxidase; O2⋅−, superoxide anion; PKG1α, protein kinase G 1α; PLA2, phospholipase A2; PLs, phospholipids; TRPV4, transient receptor potential vanilloid channel 4. Adapted from Durand et al. [
Previous studies showed the sensitivity to these pathways of vasodilation increases with decreasing vessel diameter thus assuming a particularly important role in the coronary microcirculation [19]. Previous studies have also suggested the relative weight of these pathways changes from childhood to adulthood and between healthy and pathological conditions. In a preliminary study with human-isolated arterioles, Zinkevich et al. [68] proposed the flow-mediated dilation (FMD) in infants was exclusively COX-dependent, that is mediated by prostaglandins, while in adulthood the main pathway involved the NO. However, in the presence of coronary artery disease (CAD), both these mechanisms seem to remain as secondary pathways as the EDHF-mediated vasodilation (especially by H2O2) gains importance, serving as backup mechanisms in disease [66]. In fact, low response to shear forces and high mechanical stress seem to predispose to vascular dysfunction and disease [63].
\nThe heart is a highly organized organ where several cells may be found, namely endothelial cells and cardiomyocytes. Therefore, the physiological mechanisms depend on the communication between the several types of cells. Until today, many endothelial-derived cardio-active factors have been identified and characterized (Figure 7). The cardiac modulator effects of some of these factors, such as NO, PGI2, ET-1 and neuregulin-1 (NRG-1), have been previously acknowledged. Other factors, namely Dickkopf-3 (DKK3), periostin, thrombospondin-1 (TSP-1), follistatin (FST), apelin and connective tissue growth factor (CTGF), also appear to modulate the cardiomyocyte function, though with little evidence so far. These cardio-active factors seem to be interdependent (additive, synergistic or inhibitory) as their modulator effects may be exerted on the same target cell [69].
\nCommunication between endothelial cells and cardiomyocytes. Abbreviations: CTGF, connective tissue growth factor; DKK3, Dickkopf-3; ET-1, endothelin; FST, follistatin; NO, nitric oxide; NRG-1, neuregulin-1; PGI-2, prostacyclin; TSP-1, thrombospondin. Adapted from Lim et al. [
The previously explored pathways are nowadays considered the major pathways of regulation of the vessel tone. However, other mechanisms may also come into play, such as the autonomic nervous system and circulating factors.
\nThe innervation of the coronary circulation by the sympathetic and the parasympathetic divisions of the
Moreover, several
As previously discussed, the coronary microcirculation plays a key role in the myocardial perfusion. Therefore, the presence of functional and/or structural abnormalities of this circulatory pathway may impair the myocardial perfusion and be involved alone as the main mechanism of myocardial ischaemia. These abnormalities are normally designated as
CMD type | \nClinical setting | \nPathogenic mechanisms | \n
---|---|---|
In the absence of myocardial disease or obstructive CAD | \nCardiovascular risk factors (e.g. ageing, arterial hypertension, smoking, diabetes) Microvascular angina | \nEndothelial dysfunction VSMC dysfunction Vascular wall remodeling | \n
In the presence of myocardial disease | \nCardiomyopathies (e.g. HCM, DCM) Aortic stenosis | \nVascular wall remodeling VSMC dysfunction Extramural compression Luminal obstruction | \n
In the presence of obstructive CAD | \nAcute coronary syndrome AMI | \nEndothelial dysfunction VSMC dysfunction Luminal obstruction | \n
Iatrogenic microembolization | \nCoronary reperfusion procedures (e.g. PCI) Revascularization (i.e. CABG) | \nLuminal obstruction Autonomic dysfunction | \n
Classification of CMD according to the involvement of pathogenic mechanisms and the clinical setting.
Abbreviations: AMI, acute myocardial infarction; CABG, coronary artery bypass grafting; DCM, dilated cardiomyopathy; HCM, hypertrophic cardiomyopathy; PCI, percutaneous coronary intervention; VSMC, vascular smooth muscle cell. Adapted from Crea et al. [20].
CMD may present several pathogenic underlying mechanisms, depending on the source of the abnormality, namely structural and functional (Table 1), which will be discussed in this section. According to the underlying clinical setting, the CMD may also be classified into four types: type 1, in the absence of cardiomyopathies or obstructive CAD; type 2, in the presence of cardiomyopathies; type 3, in the presence of CAD; and type 4, iatrogenic [3, 20, 73, 74].
\nThe most common functional abnormalities are the
As presented in Table 1, the traditional cardiovascular risk factors (i.e. ageing, gender, obesity, smoking, hypertension, dyslipidaemia and diabetes) may impair the endothelial function by several mechanisms, namely increased production of EDCFs and/or decreased production of EDRFs [3, 73]. Furthermore, this impairment may also contribute to the dysfunction of the VSMCs, which may also result from structural changes, derived from cardiomyopathies or arterial hypertension, described further below.
\nSimilarly to smoking,
The vascular effects of
The impairment of the vasodilator response of the coronary microcirculation may also be present in patients with angina-like chest pain but without evidence of obstructive CAD or myocardial disease. This situation is usually known as
The autonomic nervous system dysfunction, following acute myocardial infarction (AMI) and/or coronary reperfusion procedures, may also contribute to the CMD. In fact, increased coronary vasoconstriction has been previously shown, after AMI and successful percutaneous coronary angioplasty, both at the site of stenosis and distal to it, suggesting abnormal coronary vasodilator response [73, 100].
\nAfter AMI, the CMD may result from autonomic dysfunction and luminal obstruction, discussed further below. The autonomic dysfunction in the AMI-associated CMD involves increased sympathetic activation with increased vasoconstriction. These findings were confirmed by Gregorini et al. [101], who showed this impaired autonomic function might be reverted with α-blockers, such as phentolamine (nonselective α-blocker) and urapidil (α1-selective blocker), which may improve the recovery of myocardial perfusion after coronary stenting in patients with AMI [101–103]. Autonomic dysfunction secondary to percutaneous coronary angioplasty was also showed by Gregorini et al. [104] who linked the left ventricular macro- and microcirculatory dysfunction in patients with transient ischaemia. In this study, phentolamine and urapidil were similarly used to block the α-adrenergic neurotransmission and propranolol (nonselective β-blocker) and the β-adrenergic neurotransmission, and the results showed that the increased coronary vasoconstriction, secondary to percutaneous coronary angioplasty, may be prevented with α-adrenergic receptor antagonists as no effect was demonstrated for the β-adrenergic blockade. Moreover, this study suggested that CFR may still be decreased for 7 days to 3 months after the procedure [104]. In a similar study, Kozàkovà et al. [105] confirmed the potential usefulness of urapidil to improve the left ventricular function in the angioplasty follow-up. Moreover, persistent yet reversible CMD after coronary revascularization was also previously showed [106].
\nIn addition to functional abnormalities, structural abnormalities, namely vascular remodelling, vascular rarefaction, perivascular fibrosis, luminal obstruction and infiltration of the myocardium and vascular wall, may also contribute to CMD [3, 73].
\nThe remodelling of the vessel wall involves persistent modifications which may result from several
In addition to the functional changes,
Other situations may also contribute to the vascular remodelling, especially arterial hypertension and cardiomyopathies. Previous studies have suggested that
Furthermore,
The coronary microvascular function may also be influenced by modifications in the vascular density, particularly by vascular rarefaction, that is the reduction in the number of microcirculatory vessels, which may also be recognized as hypotrophic remodelling [19]. The presence of arterial hypertension and the extravascular compression, observed in aortic stenosis and cardiomyopathies, induces vascular rarefaction leading to the reduction in the CFR. In addition to the vascular rarefaction, both situations may also induce perivascular fibrosis promoting structural modifications of the vessel wall [73, 114].
\nCMD may also be characterized by luminal obstruction originated from (a) obstructive CAD or (b) iatrogenic microembolization [73].
\nAccording to the mechanisms underlying the associated CMD as well as the clinical findings, the
In patients with
Similarly to stable CAD,
Luminal obstruction is a key characteristic of the
Role of pericytes in healthy and ischaemic microvessels: (a) normal blood flow in coronary arterioles and capillaries covered with pericytes; (b) in ischaemia, the pericytes may constrict the coronary microcirculation compromising the coronary blood flow and leading to coronary microvascular dysfunction; (c) after reperfusion procedures, the coronary microvascular dysfunction may block the re-establishment of the normal blood flow, situation usually known as no-reflow phenomenon.
As previously mentioned, CMD may also be originated from
In addition to the previous explored structural changes, the infiltration of the vessel wall with metabolic deposits may also be found. This infiltration is commonly found in infiltrative diseases, such as Anderson-Fabry disease and other metabolic disorders. The Anderson-Fabry disease involves a genetically linked (X chromosome) deficiency of lysosomal α-galactosidase A, which leads to damages in several organs, namely the heart, through the deposition of glycosphingolipid in cardiomyocytes and in the vascular wall [73]. In turn, this infiltration promotes the hypertrophy and fibrosis of cardiomyocytes as well as CMD and perivascular fibrosis [73]. In fact, Elliott et al. [122] demonstrated these patients present a marked decrease in CFR, confirming the presence of CMD in the pathogenesis of the cardiomyopathy induced by this disease.
\nThis review provides an insight on the morphology, physiology and pathophysiology of the cardiac microcirculation. As discussed, the heart is one of the most nutrient and oxygen demanding organs as this demand needs to be satisfied with an adequate vascularization of the myocardium through an extensive macro- and microvascular network. Although most of the cardiac diseases, such as the acute coronary syndrome, are commonly associated with the coronary macrocirculation (i.e. epicardial coronary arteries), the coronary microcirculation also seems to play a key role in the coronary pathophysiology. This role involves both molecular and clinical aspects that should not be overlooked and that constitute potential diagnostic and therapeutic targets, particularly important in the early pathogenesis of these diseases.
\nScanning Tunneling Microscopy (STM) and Atomic Force Microscopy (AFM) metrology as parts of more general Scanning Probe Microscopy has been around for a long time, and especially intense since it has been awarded by the Nobel Prize in Physics in 1986. In 1988, our team was one the first who designed, manufactured, and used the specialized scanning tunneling microscope to measure newly developed big-size surface-relief holographic diffraction gratings obtained in non-organic photoresist and having in an order lower roughness (Figure 1) [1]. For today, STM & AFM profile measurements on surface-relief diffraction gratings are presented as a matter of routine, see, e.g., in Refs [2, 3]. On the other side, precise microscopic surface-relief patterns are used as grating standards to calibrate atomic force microscopes (see, i.e., in [4]).
STM images of holographic relief grating surfaces (Au-coated) obtained by (a) organic and (b) non-organic photoresists (after [
However, a wide analysis of the use of the STM and AFM methods for surface relief grating groove metrology has not really been undertaken in details. The following problems are discussed here: the tip deconvolution, geometry, and radius; groove shapes and abrupt groove slopes; roughness; PSD functions; other. The author demonstrates examples of AFM & STM data and comparisons with other widely-used metrology techniques for bulk, coated, and multilayer-coated ruled, or holographic, or laser-lithographic, or electron-lithographic gratings having lamellar, or sinusoidal, or blazed, or other realistic groove profiles. These gratings were chosen because high quality efficiency data exists, in particular, for flight gratings or/and X-ray gratings characterized by synchrotron radiation sources; and their groove profiles, together with random nanoroughness, were measured by AFM or STM to be included in rigorous efficiency and scattered light intensity calculus.
Here the author briefly compares various standard techniques for exact determining the digital profile and 3D topography of a surface relief grating. Several widely used direct (or semi-direct – ‘imaging’) methods and respective instruments applicable for this purpose are compared. The advantage of using direct and exact groove metrology to predict efficiency and polarization characteristics of gratings now is well-known and widely used. The main purpose of such modern approach is rejecting unusable samples on earlier stages and decreasing expenses for their production and research. This is much more effective in compare to the earlier approaches, wherein: (1) a master grating is fabricated, whether by mechanical burnishing with a ruling engine, or holographic writing (interferometry), or direct laser recording (DWL), or various newer writing techniques, like as electron-beam lithography (EBL) and Si-etching, or their combinations; then, (2) it is replicated or/and coated, and, finally, (3) tested for the diffraction efficiency and scattering light intensity. For mechanically ruled gratings, a ‘test’ ruling can quickly be checked with this approach, whereas a complete ruling sometimes requires several days or even weeks of continuous use of expensive ruling machines [5]. Even for holographic or EBL gratings, considerable efforts of writing, etching and coating the grating with specialized coatings, especially multilayer coatings, can be decreased additionally if metrology validates that an intermediate product is suitable in the planned application.
The author briefly discusses and compares in the first part of this chapter several basic, among many others, direct metrology techniques: microinterferometry (as one of optical methods) [6], stylus (mechanical) profilometry [7], scanning electron microscopy (SEM) [8], and AFM [9]. Several examples of groove metrology results are presented and discussed further including those obtained very recently.
Any method for measuring the profile of a surface relief grating requires some calibration procedure [10]. The considered here methods are also widely used for surface microroughness determination on a nanometer or sub-nanometer level. Measuring main groove parameters of a grating, in particular, the actual groove depth or blaze angle, adds to the requirements for the specific metrology method. The depth of the groove profile, defined depending on the accuracy of a vertical calibration, basically determine the wavelength for the peak efficiency in a given optical mounting geometry (classical or conical) [11]. The common error of the order of a few percent in the lateral calibration can affect the prediction of the blaze wavelength that should be within hard tolerance for many practical applications. This is because the groove vertical geometry is often expressed relatively to the grating period, in dimensionless units. Any lateral error becomes vertical error in the respective topographical transformation. Fortunately, lateral errors can be fairly determined because the grating period is well known beforehand with high accuracy and, thus, the grating data itself gives a calibration factor to correct the lateral scale unit. The accurate lateral calibration is also required for rigorous efficiency modeling codes, in which the use of the average groove profile shape is very important to obtain exact efficiency data in all significant diffraction orders.
The microinterferometer is sometimes called as ‘optical profilometer’. It is essentially an interferometric head on a microscope, where the reference arm of the interferometer views a small, highly polished reference plate [6]. Such a reference can be removed from the results of measurements on highly polished surfaces that is important for grating measurements because many state-of-the-art gratings, especially for X-ray and Extreme Ultraviolet (EUV) applications, have the root-mean-square (RMS) roughness of the same order as the best reference plates. A Phase Shift Instruments model MicroXAM [12] has been used in the discussed work [10]. It has variable magnification from 2× to 100×; values of range and resolution for the 50× magnification is listed in Table 1. The instrument uses the zero path difference calculations independently for each pixel from a series of images obtained during a vertical sweep. This increases the available vertical range and the available slope angle range substantially, however any microinterferometer has two lateral resolution-restricted factors, which are not limitations in the other considered methods. Namely, the optical resolution is due to the diffraction limit and pixel sampling is due to different magnifications and focusing. The theoretical limit on lateral resolution in such instruments is a half of the working wavelength, or, typically, about 0.1–0.3 μm. Thus, this method generally is not suitable to measure high-frequency (short-period) and/or low-depth diffraction grating.
Instrument | Microinterferometer 50× | Stylus profilometer | Atomic-force microscope | Units |
---|---|---|---|---|
Vertical resolution | 0.05 | 0.1 | 0.05 | nm |
Vertical range | 100 | 130 | ∼5 | μm |
Lateral resolution | ∼0.3 | ∼0.3 | 0.015 | μm |
Lateral range | 163 (more w/stitching) | > 25000 | 100 | μm |
Limiting factor(s) for lateral resolution | MTF, sampling, need for retroreflection over the whole profile | Tip radius & angle | Tip radius | |
Upper slope limit | – | 45 | ∼70 | deg. |
Measurement characteristics of three surface profiling instruments (after [10]).
The stylus profilometer has a diamond tip to brought into direct contact with the surface, with calibrated contact force. As the tip moves across the surface, the motion of the tip is amplified, filtered, and detected. The basic limits inherent to such metrology devices are well-discussed, e.g., in [7]. Care must be exercised to prevent indentations of the surface by the tip, depending on materials and forces used, as well as accounting the tip size. The model used in this work is a Tencor P-10 [13]. Table 1 presents the basic lateral and vertical ranges and resolutions typical for the instrument. Typical measurement parameters are: the tip radius of 0.1 μm (in the plane of dispersion), the tip speed of 5 μm/s, the digital sampling of 2 kHz, the tip force of 0.25 mg, and profile lengths of at least 100 μm (depending on the groove period). Note that in the last model of this instrument, KLA-Tencor HRP-260, the tip radius can be up to 25 nm and it has a high-resolution stage that produces scanning results similar to an AFM device. In the recent investigation we have used another model, namely, ХР-1 Stylus (Ambios Тechnology, USA) [14].
The AFM instrument model that has been used in the discussed earlier work was a Digital Instruments Nanoscope III [15]. The recent investigation was carried out using the atomic-force microscope model NT-MDT NTEGRA Aura [16]. NTEGRA Aura is a Scanning Probe Microscope for studies in the conditions of controlled environment and low vacuum. The Q-factor of the cantilever in vacuum increases, thus gaining the sensitivity, reliability and accuracy of ‘probe-sample’ light forces measurements. At that, the change from atmosphere pressure to 10−3 Torr vacuum provides the tenfold gain of Q-factor. By further vacuum pumping, Q-factor reaches its plateau and changes insignificantly. Thus, NTEGRA Aura comparing to the high-vacuum devices it needs much less time, about a minute, to get the vacuum that is needed for the tenfold Q-factor increase. NTEGRA Aura has built-in closed loop control for all the axes, optical system with 1 μm resolution and ability to work with more than 40 different AFM regimes.
We have used for the Si-grating technology investigation a flexible Carl Zeiss SUPRA 25 SEM system with a versatile analytical specimen chamber that can be easily expanded with a choice of optional detectors and a full range of accessories [17]. Utilizing the unique GEMINI field emission column, it delivers superb resolution over the complete high voltage range with the magnification of up to 500000. The large 5-axes motorized cartesian stage is particularly useful for handling a number of smaller specimens simultaneously. It is equally useful for accommodating bulky or irregular shaped specimens.
Table 1 summarizes the capabilities and limits of the three metrology devices, which have been characterized earlier for grating metrology. As one can see, the atomic-force microscope has the finest lateral and perfect vertical resolutions. The stylus profilometer and the microinterferometer have comparable vertical ranges, however, without a possibility to determine superfine (atomic-scale) structures, i.e., nanoroughness, and abrupt slopes (see, e.g., Figures 3–5). On the other hand, the stylus profilometer has significantly larger lateral range for probing to the millimeter spatial range. Also, the AFM data gives a typical example of non-linearity that should be accounted and described further.
In the groove profile experiment, a series of step height calibration standards [18] has been used [10]. The vertical axis was calibrated using one of the smallest steps of 10 nm. Then the rest of the step height series was measured. Small errors, up to 8%, were observed for heights much higher than that used to calibrate the atomic-force microscope. The fit to correct such nonlinearity was used when the nonlinearity gives a significant difference. The obtained results are summarized in Table 2.
Nominal height, nm | Microinterferometer | Stylus profilometer | Atomic-force microscope |
---|---|---|---|
8.7 | 7.92 | 8.1 | — |
25.8 | — | — | 25.7 |
42.7 | 43.4 | 42.4 | — |
530 | — | — | 520 |
1046 | — | — | 1005 |
1590 | — | — | 1469 |
960 | — | — | — |
Note No. | 3, 4 | 2 | 1 |
Step height data (after [10]).
1. Nonlinear at ∼8% at highest step when calibrated to a 10 nm step.
2. Using 0.1 μm tip, could not resolve depth of 3.3 μm period, AFM step height standard.
3. Used at 50× magnification.
4. At 100× did not have lateral resolution to see the 3 μm period samples tested using AFM.
In the manufacture and analysis of diffraction gratings, it is necessary to control certain of their parameters at each stage of the process. A SEM research [19] is permissible only at the stage of development of the manufacturing technology, because after each technological operation, see, e.g., [20] or Section 4.5, the sample of the Si-etched grating becomes less and less, since a fragment is separated from the sample to obtain a SEM image of a transverse cleavage (CS). In contrast to this, AFM studies are non-destructive; therefore, control of parameters in the manufacture of gratings is usually carried out with the help of AFM. We made a comparison between AFM (NTEGRA Aura microscope) topographies and SEM (SUPRA 25 system) images of Si-etched grating samples with the period of 2 μm. Table 3 shows the results of AFM and SEM investigations of Si-etched gratings obtained at different stages of their fabrication. Table also presents the numerical comparing between the AFM and SEM results of the measured groove geometric parameters for the samples studied. The calculated value of the blaze (working facet) angle from the SEM studies was obtained from the sine determined by dividing the experimental values of the groove depth by the width of the working facet.
Sample No. | Groove depth, nm | Working facet width, nm | Working facet angle, deg. | |||
---|---|---|---|---|---|---|
SEM | АFM | SEM | АFM | SEM | АFM | |
1 | 151 (47)* | 133 (38)* | 1571 | — | — | — |
2 | 149 (37)* | 141 (44)* | — | — | — | — |
3 | 111 | 121 | 1630 | 1710 | 3.90 | 4.05 |
4 | 111 | 111 | 1603 | 1594 | 3.97 | — |
5 | 105 | 114 | 1590 | 1580 | 3.89 | 4.13 |
AFM and SEM data for blaze Si-etched gratings.
Height of Si-nubs.
To measure the roughness of Si(100) plates etched through a DWL mask in KOH and intended for developing the technology of manufacturing Si-etched gratings, several high quality samples were selected using white light optical microscopy. The roughness of the etched bottom and the non-etched area was measured by two compared methods: Stylus Profilometry (XP-1 Stylus profilometer) and AFM (NTEGRA Aura microscope). Our studies were carried out on an atomic force microscope in the semi-contact or tapping mode; all scans had 512 × 512 points. We used TipsNano [4] silicon cantilevers with a typical radius of tips ∼6 nm. The results of roughness measurement by two methods on topological elements (stripes) of 50 μm wide are presented in Table 4. As follows from the presented data, the RMS roughness obtained by different methods may differ by more than an order of magnitude. This is due to the radius of the stylus and the scanning length, which in that case were 2 μm and 80 μm, respectively. However, this device is equipped with a stylus with a radius of 0.2 μm, which, in principle, allows one to measure low- and mid-frequency gratings with smaller roughness. Note that for the etched bottom, where the average roughness is several times higher, the scatter of results is much smaller and ranges from several tens of percent to several times.
Sample No. | RMS roughness, nm | |||
---|---|---|---|---|
Non-etched field | Etched bottom | |||
Profilometer | AFM | Profilometer | AFM | |
1 | 2.6 | 0.2 | 2.8 | 0.8 |
2 | 3.2 | 1.6 | 4.0 | 2.5 |
3 | 3.4 | 0.2 | 4.5 | 1.8 |
4 | 2.8 | 1.7 | 2.9 | 4.9 |
5 | 2.4 | 1.5 | 3.5 | 4.3 |
AFM and stylus Profilometry data for Si-etched plates.
A cantilever tip convolution, which limits the resolution of both the atomic-force microscope and the stylus profilometer, has been much studied and various algorithms to account for this effect has been developed and intensively used (see, e.g., [21] and also in this book). In the results presented in Table 2 such algorithms have not be used. However, the general used rule is that the known tip radius should be much less than the measured periods of gratings. Typically, the radius of a fresh AFM cantilever tip is about 5–15 nm; so, the rule of thumb is that for groove profiles of mid- and high-frequency gratings (say, periods of 100–300 nm and less) tip deconvolution algorithms should be used. In the vertical direction, the depth parameter is smaller, and, apparently, the groove profiles recorded somewhat non-correctly for high-frequency gratings only. However, it depends also on absolute values of the groove profile depth, which can vary in two orders of magnitude.
Another important and general AFM problem, in particular for fine-structure gratings with steep slopes and high aspect ratios of grooves, is the shape and the radius of AFM cantilever tips. Tip size has the major impact on the resolution of images obtained by any atomic-force microscope. The knowledge of the tip radius and shape is essential for the quantitative interpretation of nano-scale lateral steps, in particular, for roughness having short correlation lengths. Tip wear is therefore a key limitation in the application of AFM [22]. The results of nanoindentation experiments with diffraction gratings permanently confirm this conclusion. The measurement of the tip radius before and after measuring groove profiles of gratings was performed in Ref. [10], and they found that the radius to be in the range of 10–20 nm. One measurement found a fresh tip to be ∼10 nm radius and a used one to be ∼20 nm. Therefore, one should restrict an AFM-profiling work to gratings of period much longer than 10–20 nm, as it has been discussed above.
One more problem in AFM measurements of diffraction gratings is the grooves with steep facet slopes, which can be 80 degrees and more for echelle gratings [2]. This problem is similar to measurements of the rectangular (lamellar) groove profile in microelectronics [23]. To accurately measure such general trapezoidal profiles with steep or even negative sidewalls, a large change in the angle of inclination of a cantilever (or scanner, or sample) and/or special cantilever (tip) shape are required, as well as taking into account the aspect ratio of measured grooves [24]. Several studies applied to periodic structures demonstrate that some combination of the tilted probe, special orientations of AFM images and appropriate deconvolution algorithms allows the precise groove shape reconstruction at any aspect ratio [25]. An example of such problem successfully solved is the average groove profile (two grooves) of a 112/mm echelle R5 grating (blaze angle ∼78°) derived from AFM images and presented in Figure 2.
Average AFM groove profile for 112/mm echelle R5 grating.
In Figure 3, typical power spectral density (PSD) 1D functions for Si(111) substrate and Si-etched grating samples (see also Section 4.5) are shown. An estimator of the PSD function is factually the periodogram for any periodic, or quasiperiodic, or random profile, or some combination. Assuming the ergodicity of a stochastic process connected with a random generation of asperities, the PSD function can be found as the Fourier transform of the autocorrelation function [26]. Although these functions are mathematically equivalent, one can analyze easy any corrugations of the profile shape simultaneously, i.e., random roughness and groove depth variations, using the 1D or 2D PSD function. Then, the RMS roughness is directly calculated through PSD as the root square of the integral over an effective range of allowed spatial frequencies. Thus, a wide lateral scanning range may require for an AFM instrument to take into account in the evaluated RMS roughness all spatial frequencies (or correlation lengths). It is especially important for low-frequency (long-period) gratings having additionally large correlation lengths of random roughnesses. A good discussion related to this problem and devoted of the use of AFM and similar instruments for measurements of PSD functions of smooth mirrors for imaging systems working in the X-ray–EUV range can be found in [27]. So, if one need to use images with a scanning area of about 100 × 100 μm2 then thermal drifts, hysteresis, and essential scanner nonlinearities should be accounted.
PSD function: (left) for Si(111) substrate; (right) for Si-etched grating with 500/mm and 4° blaze angle.
The abovementioned metrology techniques were applied to validate the efficiency of a chosen grating from an ordered grating set which is mounted in the Space Telescope Imaging Spectrograph (STIS) flown aboard the Hubble Space Telescope (HST) [28]. A − 1-order reflection grating with 67.556/mm blazed for 750 nm (1.44° nominal blaze angle) working in the range from 500 to 1000 nm at 8° incidence angle was chosen by us for a certification [29]. The pattern size was 1.5 inches by 1.5 inches, and the ruled area was 30 mm by 30 mm. A sister-replica to this grating, designated ‘Ng41M’ or by its manufacturers’ (Richardson Gratings of Newport Corp.) serial number, 1528, is in use on the HST/STIS as a red survey grating (blazed in the red visible and near infrared range) [10]. In its flight application, this grating had a reflective overcoating of 100 nm Al plus 25 nm MgF2. However, in these wavelengths the effect of the MgF2 layer is minor and simulations have showed no valuable difference, within a small part of the accuracy in the measured diffraction efficiency) with such coating or without it. This grating was chosen as an example because: (1) high quality efficiency data exists for it, including rigorous efficiency calculus using the realistic groove profile shape; and (2) groove profiles can be measurable by the mentioned above three methods for a direct comparison.
Portion of a trace of grating No. 1528 taken with the microinterferometer is shown in Figure 4. Both the depth and the profile shape are somewhat distorted in compare with the groove profiles in Figures 5,6 obtained by the other considered methods. However, as one can see, the overall groove depth and profile are evident. It is clear from the all figures that the profile roughness is higher on the upper sloped portion than on the steep edges. Difficulties in holding the sample steady during ‘flyback’ prevented reproducibility of measurements for that microinterferometric study.
Portion of a microinterferometer trace of ruled grating No. 1528 (after [
Portion of a stylus profilometer trace of ruled grating No. 1528 (after [
AFM image of two grooves of ruled grating No. 1528 (after [
The groove profile was characterized in details AFM measurements. The tips used here were 10 or 20 nm in radius. An example of the typical groove profile of No. 1528 grating is presented in Figure 6. Figure 6 shows an example of AFM data for a portion of the surface of the investigated ruled grating. The basic groove profile shape is clearly evident, along with portions of the profile that are rougher than others, and some roughness along the grooves is indicated as well. Figure 6 shows that the minimum of the grooves is clearly visible in the AFM image. If, as usually, one selects the bottom of the groove as the minimum value, there are two complete grooves in each scan.
The resulting average groove profile – with averaging performed both across the grooves and along as well – is shown in Figure 7. The solid line is based on the AFM data, and the dotted line is based on the stylus profilometer data: the groove tops are aligned for the purpose of this comparison; the relatively sharp groove bottom is not as well resolved by the stylus profilometer. The periodicity of the profile is shown by comparing a model of the averaged scan based on the average groove profile shape to the average scan. This is demonstrated by dotted lines plotted against the initial data in Figure 4 (microinterferometer) and Figure 5 (stylus profilometer). Once the average profile has been determined, the fitting routine finds the sawtooth and two-angle shape fits by the method of least squares. It is found in the considered case the blaze angle of 1.45° and the anti-blaze angle of 30° (Figure 8). The efficiency in general is fairly insensitive to the anti-blaze angle, and the fitting procedure does not fit it as consistently as it does a case of the blaze angle. Thus, the final average groove profile derived from AFM measurements for efficiency modeling purposes is shown in Figure 8 with 100 discretization points [29].
Average groove profile for grating No. 1528 based on AFM and stylus profilometer data.
Models of normalized to period groove profiles of No. 1528 grating.
The surface of gratings, namely, the master [30] and replica [31] gratings, as examples of ‘good’ products, were characterized using a Topometrix Explorer Scanning Probe microscope [32], a type of atomic-force microscopes. The gratings have 2400/mm, a concave radius of curvature of 2.0 m, and a patterned area of size 45 mm by 35 mm. The master grating was fabricated by Spectrogon UK Limited (formerly Tayside Optical Technology). The groove pattern was developed in fused silica by a holographic technique using ion-beam etching to produce an approximately triangular, blazed groove profile. Ion-beam etching results in a groove profile much closer to triangular than the ideal blazed (sawtooth) profile with the apex angle of ∼90°. The master grating was uncoated. The replica of the master grating was produced by Hyperfine, Inc. As a result of the replication process, the replica grating had an aluminum surface. A thin SiO2 coating was applied to the Al surface for the purpose of reducing the nanoroughness and protecting the surface from an additional oxidation.
The AFM images typically had 500 × 500 pixels and a scan range of 1 to 20 μm (pixel size 20 to 400 Å). The silicon probe had a pyramid shape. The base of the pyramid was 3 to 6 μm in size, the height of the pyramid was 10 to 20 μm, and the height to base ratio was approximately 3. The tip of the pyramid had a radius of curvature 100 to 200 Å. The AFM scans were performed using the non-contact resonating mode, where the change in the oscillation amplitude of the probe is sensed by the instrument. A surface topology reference sample was used to optimize the AFM scanning parameters, to calibrate the height scaling of the instrument, and to evaluate the performance of the AFM. This was essential for the accurate characterization of the gratings. The surface topology reference sample consisted of an array of approximately square holes fabricated on the silicon dioxide surface of a silicon die by VLSI Standards, Inc. [18]. The top surface of the die was coated with a thin layer of Pt. The hole array had a pitch of 3 μm and a hole depth of 180 Å.
One typical AFM image of the master grating measured using 16-Å pixels is shown in Figure 9, where the vertical scale has been scaled to reveal the texture of the groove surface. The RMS roughness, determined by integrating the PSD function over 2–40 μm−1 range, was 3.2 Å. Most of the roughness is concentrated at low spatial frequencies as is apparent from the analysis of the PSD function. The central portion of the AFM image shown in Figure 9 that covers one period of the grating pattern was selected for further investigations. An analysis program was written in the Interactive Display Language (IDL) for this purpose and it is discussed in detail in Ref. [31].
AFM image of 2400/mm holographic (master) grating (after [
The histogram of the pixel heights, for one period of the grating pattern, is shown in Figure 10. The maxima at 10 Å and 85 Å in Figure 10 are caused by rounding of the groove profile at the peaks and the troughs which is a result of the pattern fabrication process. An ideal groove profile, either sawtooth or triangular, would have a flat height histogram. The separation between the peaks in Figure 10 represents the average groove height, approximately 75 Å. The local blaze angle at each pixel was determined by using a least squares algorithm to fit a linear curve to the data points in a sliding window. The window was 25 pixels (400 Å) long in the direction perpendicular to the grooves and one pixel wide parallel to the grooves. The blaze angle is the arctangent of the fitted slope. The histogram of the blaze angles, for all rows of data in one period of the grating, is shown in Figure 11. The peak at 2.5 deg. represents the classical blaze angle, and the peak at 5.5 deg. represents the steep facet of the ideal sawtooth profile as modified by the ion-beam etching process. For a density of 2400/mm and for facet angles of 2.5 deg. and 5.5 deg., an ideal grating would have a groove height of 125 Å. However, the measured value of 75 Å (Figure 10) indicates a significant degree of rounding at the peaks and troughs of the groove profile. In addition, the measured ratio of the heights of the 2.5 deg. and 5.5 deg. features in the angle histogram (Figure 11) is approximately 3, greater than the ratio of approximately 2 that is expected based on the average facet angles.
Histogram of pixel heights from AFM image of 2400/mm holographic grating (after [
Histogram of blaze angles from AFM image of 2400/mm holographic grating (after [
The interpretation of the widths of the features in Figure 11 is difficult because they are complicated functions of the surface roughness, the width of the sliding window, and the probe geometry. This is addressed in the publications [30, 33, 34]. The feature at −2 deg. in Figure 11 results from the fits to the peaks and troughs of the groove profile, where the local slope is changing rapidly but has an average value near zero. Simulations show that the −2-deg offset of this feature from zero is a consequence of the unequal average blaze angles of the two facets. To provide a groove profile for the efficiency calculation, a representative AFM scan perpendicular to the grooves was chosen at random and scaled to the average groove height. The resulting groove profile is shown in Figure 12. This groove profile has 210 points.
Average groove profile from AFM image of 2400/mm grating: (1) 7.5-nm- deep master; (2) 9.0-nm-deep replica; (3) 6.6-nm-deep scaled replica (after [
AFM image of 2400/mm replica grating (after [
An AFM image of two grooves of the replica grating is shown in Figure 13. The scan was performed across the grooves over a range of 1 μm (20-Å pixels). The vertical scale in Figure 13 has been expanded to reveal the texture the texture of the grating surface. The PSD function derived from a 2 μm-size image spanning nearly 5 grooves is shown in Figure 14. The peak in the 2 to 3 μm−1 frequency range results from the 0.4167 μm groove period. The RMS roughness is 7 Å in the 4–40 μm−1 frequency range. By comparison, the RMS roughness of the master grating measured by the same type of AFM instrument was 3.2 Å, and this implies that the replica grating is significantly rougher than the master grating. This may result from the replication process, which for a concave grating is at least a two-step process. Furthermore, the master grating was fabricated on a fused silica surface by a holographic technique and was ion-beam polished, while the Al surface of the replica grating may contribute to its larger nanoroughness. The replica grating without the SiO2 coating was not characterized by AFM. Typical average groove profile derived from the AFM image (1 μm in size) of the replica grating is shown in Figure 15. The groove profile is approximately triangular in shape with rounded corners and troughs and with facet angles of 3.4 deg. and 6.2 deg. The average groove depths derived from the AFM images are in the range 85 to 95 Å. These values of the facet angles and the groove depth are larger than the corresponding values for the master grating, 2.5 deg. and 5.5 deg. facet angles and 75 Å average groove depth (Figure 12). Thus, the grooves of the replica grating are deeper and the facet angles are steeper compared to those of the master grating.
PSD function of 2400/mm replica grating from AFM image (after [
Average groove profile from AFM image of 2400/mm Mo4Ru6/Be grating (after [
Multilayer gratings were produced by application of Mo4Ru6/Be multilayer coatings [35, 36] to two replicas of the described holographic master grating. Beryllium-based multilayer coatings can provide substantial reflectance at wavelengths near 11 nm. Such a Mo4Ru6/Be multilayer coating with 50 bi-layers was applied to the grating substrate. The coating was deposited by the magnetron-sputtering technique. Here we describe one of the multilayer gratings.
The surface of the multilayer grating was also characterized using the same Topometrix Explorer scanning probe microscope. The grating topography was measured merely for the master, replica, and multilayer gratings. The scan was performed across the grooves over a range of 1 μm (2-nm pixels). Typical groove profiles derived from the AFM image (1 μm in size) of the master, replica, and multilayer gratings are shown in Figures 9,13,15, respectively. These groove profiles have from 120 to 210 points. The groove profiles are approximately triangular in shape with rounded corners and troughs and with facet angles of 2.5° & 5.5°, 3.4° & 6.2°, and 3.0° & 4.1°, respectively. The average groove depths derived from the AFM images are in the range 7 to 8 nm, 8.5 to 9.5 nm, and 8 to 9 nm, respectively. Within the AFM groove-to-groove variation of the facet angles, the border shapes did not significantly change after multilayer coating. As determined above the average surface of the multilayer grating was characterized using a scaled replica AFM profile (Figure 12).
The aforementioned AFM method was applied to simulate the efficiency of a 5870/mm G185M grating intended for operation at vacuum-ultraviolet (VUV) wavelengths below 200 nm [37]. This grating has the highest groove density and the shortest operational wavelength range of all Cosmic Origins Spectrograph (COS gratings planned for the last servicing mission to the HST) [38]. The G185M master grating was recorded holographically on 40 mm by 15 mm rectangular fused silica blank and the Pt coated at HORIBA Jobin Yvon Inc. [3]. An adhesive Cr coating, a working Al coating, and a protective (from oxidation) MgF2 coating were deposited on Au-coated replica gratings at NASA/GSFC.
Resonance efficiency anomalies associated with waveguide funneling modes inside the MgF2 dielectric layer degrading the G185M COS NUV grating performance were measured and qualitatively described at NASA/GSFC [39]. We used PCGrate-SX v. 6.1 [40] to model the efficiency of the G185M subwavelength grating with real boundary profiles (measured by AFM) and refractive indices (RIs) taken from different sources, including best fits of the calculated efficiency data to experimental ones [37].
The border profiles were characterized using AFM measurements. The profile of the G185M grating (replica C) intended for operation in the 170–200-nm range was AFM-measured before and after deposition of the Cr/Al/MgF2 coating (Figure 16). As seen from the figure, after the deposition the profile depth decreased by about a factor of 2.05 (46.4 nm against 22.6 nm), and the profile shape changed noticeably too, thus evidencing the case of nonconformal layering of the grating. For the reason that all G185M gratings were manufactured from the same master and by the same technology, one may suggest that all of them share before- and after-coating profiles. The average before-coating groove profile had 165 points and the average after-coating profile had 163 points.
G185M AFM-measured surfaces before (left) and after (right) coating Al plus MgF2 (after [
To determine which of the two AFM-measured boundary profiles, MgF2 (border profile 1 measured after Cr/Al/MgF2 coating) or (Cr)-Au (border profile 2 measured before Cr/Al/MgF2 coating), is closer to the MgF2-Al boundary, we started with modeling the non-polarized (NP) efficiency of a two-boundary grating. We assume a conformal MgF2 layer (the lower MgF2-Al boundary is identical in shape to the MgF2 one) with the 40.1 nm thickness. The calculated efficiencies (Figure 17, pink curve) differ from the measured values in time throughout the whole wavelength range, thus implying invalidity of a model with a conformal layer. All calculated efficiency data presented in Figure 17 were obtained with the RIs of Al and MgF2 taken from the handbook of Palik [41]. Although hereinafter the experimental efficiency data of two grating replicas (A and B) are displayed, we will focus primarily on discussing the grating A data (solid dark blue squares in Figure 17), because replica A is the grating on which more measurements were performed.
G185M –1st-order NP efficiency measured and calculated for different layer shapes.
The next step is to use two models with nonconformal layers, one with the lower boundary being the same as border 2 (Figure 17, yellow curve) and the other with the boundary scaled from border 2 at all points by a factor of 0.488 to the profile depth of border 1 (Figure 17, bright green curve). In both cases, a vertical displacement of one boundary with respect to the other (shift of the boundary reference levels) was 40.1 nm, as in the conformal model. As evident from Figure 17, the nonconformal model with unscaled lower boundary yields a noticeably superior qualitative agreement with experimental data. This suggests that the MgF2-Al boundary more closely resembles border profile 2 than border profile 1. The model takes into account the fact that the thickness difference of 23.8 nm between the lower and upper boundaries should be added to the conformal vertical displacement (40.1 nm) to obtain an adequate vertical displacement for the nonconformal MgF2 layer. In this way the period-averaged thickness of the nonconformal MgF2 layer is kept approximately equal to 40.1 nm within the boundary shape distortion.
To determine the effect of profile shape, we set up models with equal depths and vertical shifts. The first one has border 1 scaled to the depth of border 2 (making it grater by a factor of 2.05) and a vertical displacement between the zero boundary levels equal to 63.9 nm. As seen from Figure 17, the efficiency of this model (orange curve) is close to that of another model with unscaled border 2 and a vertical shift of 63.9 nm (sky blue curve), while it is inferior by 40% or more as far as matching the experimental efficiencies. The latter suggests that, to set up an exact model, one has not only to determine the depth of the MgF2-Al boundary but also to take into account the shape of its profile – see Figure 18.
Average G185M AFM border profiles before and after coating Cr/Al/MgF2.
Having determined the type of the MgF2-Al boundary profile, we have to refine it by scaling the shape in depth and then comparing the efficiencies obtained for each model with experimental data. Another fitting parameter is the vertical displacement of the boundaries. By automatic modeling of the efficiency over a small-meshed grid of these two parameters and wavelength, one can determine the average thickness of the MgF2 layer from the best fit between the calculated and the experimental efficiencies. Even slight changes (with a few nanometers) in profile depth and vertical displacement give a noticeable rise to the efficiency at fixed wavelengths, particularly in resonance regions. Figure 17 presents an efficiency curve (heavy dark blue) for the model with a lower-boundary scaling factor of 1.04 and a vertical displacement of 68.5 nm. The model with these parameters of the layer geometry provides the better least-squares fit (not worse than 20%) of calculated efficiency to experimental data, both in the medium and in the long-wavelength ranges. As to the short-wavelength part, no variations in the lower boundary profile chosen within our approach yield theoretical values of the efficiency close enough to the measured ones.
Five-boundary G185M grating model. Horizontal and vertical scales are different (after [
What only remains is to check whether the average-thickness parameters of the MgF2 nonconformal layer used in the final model provide a better fit between the calculated and experimental values of efficiency throughout the wavelength range with a new MgF2 RI library (Keski-Kuha–Goray) [37]. To do this, we scale the vertical displacement and boundary parameters for the final model. Graphical results of this three-parameter optimization (scale, shift, and wavelength) are displayed in Figure 19. The final geometrical model of border shapes and layer thicknesses is demonstrated in Figure 18. The optimization procedure using different thicknesses for all the layers accounted has been applied using the least-square method. An analysis of these results shows that the parameters of the final model do indeed provide the best agreement between the measured and calculated values of efficiency throughout the wavelength range. The relative deviation of experiment from theory for all wavelengths at which grating A was studied does not exceed 10% throughout the wavelength range. Figure 21 presents also an efficiency curve (sky blue curve) calculated by use of the approximate values of the MgF2 absorption index; all other parameters of the final model remain intact. A comparison of the curve efficiencies based on scaled (sky blue curve) and exactly calculated (heavy dark blue curve) values of absorption shows that the efficiency changes at the wavelengths where the RI imaginary values scale only slightly are indeed appreciable.
Grazing-incidence off-plane gratings have been suggested for the International X-ray Observatory (IXO) [42]. Compared with gratings in the classical in-plane mount, X-ray gratings in the off-plane mount have the potential for superior resolution and efficiency for the IXO mission [43]. The results of efficiency calculations for such a 5000/mm gold-blazed soft-X-ray grating in a conical (off-plane) mount using the average groove profile derived from AFM measurements was presented in [44].
An AFM study of the grooved area confirmed the larger than expected blaze angle. The AFM scans across the grooves near the center of the grating are shown in Figure 20(a), where each scan is displaced vertically by 1 nm for ease of viewing. The standard deviation of the data points from the average scan curve is 0.89 nm and is a measure of the roughness of the groove profile. The histogram of the angles between each pair of scan points is shown in Figure 20(b), where a Gaussian curve is fitted to the angle distribution. The top corners of the groove profiles are rounded, and this results in a rather broad distribution of angles with a centroid value of 13°.
(a) AFM scans across the grooves near the center of the grating; (b) histogram of the angles of pairs of points on the AFM scans giving a measure of the average blaze angle (after [
The average values of the blaze angles measured at seven points distributed on the grooved area ranged from 8.9° to 15°, and the RMS roughness values ranged from 0.66 to 0.92 nm. Thus, there was considerable variation of the grooves over the 5 cm patterned area. AFM data that were taken before the titanium and gold coating of the imprinted grating showed RMS roughness of approximately 0.2 nm and blaze angles of around 8°, which indicate that deposition of the metal films onto the polymer-based imprint resist led to the observed changes in groove profile [45]. High diffraction efficiencies of the Au-imprinted 5000/mm grating using the average groove profile with 123 nodes of the polygonal groove profile derived from the AFM measurements (Figure 21) are demonstrated in Refs [44, 46].
Normalized average groove profile of an Au-imprinted 5000/mm grating measured by AFM (after [
For medium- and high-frequency diffraction gratings, classical (in-plane) diffraction gives acceptable values of the efficiency of working orders only in the soft X-ray and EUV ranges [47]. However, grazing conical (off-plane) diffraction schemes have great advantages in efficiency when such gratings operate in short-wavelength regions of the X-ray spectrum (hard X-rays and tender X-rays), including in high orders and to obtain high dispersion and resolution. With such a mount, record efficiency, close to that of a respective mirror, can be obtained for sawtooth gratings with blaze angles of several degrees, which are much easier to manufacture. For a theoretical analysis of the diffraction efficiency of such gratings, the use of rigorous electromagnetic theories is required [48, 49].
The manufacturing process of a reflective Si-etched grating of a triangular groove profile (sawtooth or blaze) can be conventionally divided into four main steps: (1) obtaining a pattern of a protective mask for etching grooves (DWL or EBL, in our case); (2) anisotropic etching of grooves in a solution of potassium hydroxide (KOH); (3) etching to smooth the grating profile and polish the surface of the reflective (working) facets; (4) coating to increase reflectivity. In turn, each step consists of several operations that should be controlled using AFM and, if possible, SEM. Some AFM results (NTEGRA Aura microscope) obtained during the grating manufacturing process are considered further in detail.
To transfer the grating pattern directly to a silicon wafer (stage 2), it is etched in KOH with various concentrations at a temperature from room temperature to 50°C with vigorous stirring of the solution [20, 50, 51]. KOH etches the {111} planes more slowly than the rest of silicon, which leads to angular facets with a facet tilt determined by the orientation of the {111} planes relative to the surface plane (i.e., vicinal Si(111) plates). Therefore, KOH etches the pattern of the grooves in the Si while simultaneously setting the blaze angle of grating facets. The author uses here the results of our original Si-etched grating production technique, however, with references to the similar methods for mastering such gratings.
In our AFM studies, the following was performed: measurement of the surface roughness of the working facet on an area of 1 × 1 and 10 × 10 μm2 and measurement of the grating profile, etching depth and blaze angle of the working facet when scanning 10 × 10 μm2. The measurements were made in the tapping mode using scans of 512 × 512 pixels. We used TipsNano [4] silicon cantilevers with a typical radius of tips ∼6 nm. Examples of the AFM topography of Si-etched grating samples with a smoothed profile on the area of 1 × 1 and 10 × 10 μm2 are shown in Figure 22a,b for sample No. 5.
Surface topography obtained by AFM scanning of area of sample No. 5: a) 1 × 1 μm2; b) 10 × 10 μm2.
Figure 23a shows the topography profile of specimen No. 5 along line 1 (black curve) and the blaze angle of the working facet (blue curve). The angle is calculated as the arctan of the coordinate derivative and converted to degrees. Figure 23b shows the profile of the slope of the non-working facet along line 1 for sample No. 3/1.
Profile topography obtained by AFM scanning of area of 10 × 10 μm: a) surface and blaze angle of the working facet, sample No. 5; b) anti-blaze angle of the non-working facet, sample No. 3/1.
The results of AFM studies of the geometrical groove parameters of the samples of Si-etched gratings with a period of 2 μm are presented in Table 5. In the results presented in Table 5 the deconvolution algorithms have been used, although we evaluated mid-frequency gratings. The histogram (normalized density of probability) of blaze, anti-blaze, and apex angles of grooves of the grating with 500/mm and 4° blaze angle is demonstrated in Figure 24 (left). The three peaks on this curve are clearly associated with the corresponding working and non-working facet angles, as well as with the angle of the smoothed top of the groove profile. The average groove profile topography and the respective angles one can see in Figure 24 (right). The peak corresponding to the blaze angle is pronounced and indicates a high quality of the developed sawtooth grating. The average groove profile derived from AFM data for one grating was used then for rigorous calculus of 3D diffraction efficiencies of orders vs. incidence angle and wavelength in the soft-X-ray–EUV range and classical mount (Figure 25). The other AFM groove profile data for similar Si-etched gratings produced by DWL, or EBL, or holographic recording can be found in [52, 53] and references there in.
Sample No. | Groove depth/Si-nub height, nm | Working/non-working facet width, nm | Working facet RMS roughness, nm | Blaze angle, deg. | Anti-blaze angle, deg. |
---|---|---|---|---|---|
1 | 95/38 | 1512/340 | 0.462 | — | — |
2 | 97/44 | 1544/340 | 0.345 | — | — |
3/1 | 121 | 1710 | 0.278 | 4.05 | 20 |
3/2 | 111 | 1594 | 0.340 | — | — |
5 | 114 | 1580 | 0.337 | 4.13 | 20 |
Groove geometrical parameters of Si-etched grating samples according to AFM.
AFM groove parameters of 500/mm and 4° blaze grating: (left) histogram of groove angles including smoothed groove top (‘transition’); (right) average groove topography and respective angles.
3D diffraction efficiency in principal orders of 500/mm Au-coated Si-etched grating rigorously calculated using the realistic groove profile vs. incidence angle and wavelength.
In order to reduce the roughness of the grating surface, the authors of [54] use a nine-cycle RCA-1/HF etching procedure to remove any irregularities and roughness, i.e., perform both smoothing and polishing etching; and they report submicron roughness. To reduce the roughness of the working facet at the polishing stage, several etchants have been tested, including tetramethylammonium hydroxide (TMAH) and the isotropic silicon etchant HF: HNO3: H2O. Table 6 shows the AFM results of processing in different etchants of the surface of samples, punctured from the same grating immediately after anisotropic etching in KOH. As one can see from Table 6, the RMS roughness of working facets can be reduced to <0.3 nm for a few etching processes. The initial RMS roughness (before a polishing process) was ∼1.2 nm (compare with results in Table 5).
Sample No. | Working facet RMS roughness, nm | Polishing etchant/etching time, s |
---|---|---|
1 | 0.269 | Isotropic/30 s |
2 | 0.244 | Isotropic/20 s |
3 | 0.271 | Isotropic, using HF before/60 s |
4 | 0.315 | Isotropic/60 s |
5 | 0.246 | TMAH/2 min |
6 | 0.291 | TMAH/4 min |
7 | 0.336 | TMAH/6 min |
8 | 0.312 | TMAH/8 min |
76KDB Si(111)4°- substrate, ∅76.2 mm | 0.149 | No process |
Groove roughness of Si-grating samples according to AFM after polishing.
In the chapter, some earlier and recent results of the use of AFM & STM methods for groove metrology of various surface relief (ruled, holographic, lithographic, imprinted) diffraction gratings, mostly intended for short wavelengths, were described and discussed. Examples of a few comparisons with the other widely-used direct metrology techniques, such as SEM, stylus profilometry and microinterferometry, were also demonstrated and compared. In addition, the most critical problems connected with AFM methods for groove metrology of bulk, thin-film-coated and multilayer-coated gratings were discussed, such as: the tip deconvolution and its radius; groove shape and abrupt groove slopes; RMS nano-roughness and PSD functions.
The detailed AFM groove metrology results were presented by the author for several important grating samples: the Space Telescope Imaging Spectrograph grating flown aboard the HST and working in the Visible–NIR; the similar master, replica and multilayer soft-X-ray–EUV blaze gratings; the Cosmic Origins Spectrograph grating used in the last servicing mission to the HST and working in the VUV–NUV; imprinted off-plane blaze grating planned for the International X-ray Observatory and working in the soft X-rays; and recently developed Si-etched blaze diffraction gratings indented to work in the X-rays–EUV at high efficiency and a very low level of scattering light. These gratings were chosen because high quality efficiency data exists, in particular, for space gratings or/and X-ray gratings characterized by synchrotron radiation sources; and their groove profiles, together with random nanoroughness, were measured by AFM to be included in rigorous efficiency and scattered light intensity calculus.
The rigorous calculation accounts for the real profile of the grooves and their thickness as well as suitable refractive indices. It was not possible earlier to achieve such good agreements between measured and calculated efficiencies of high- and mid-frequency gratings working in the short spectral ranges due to the lack of realistic, i.e., measured using the AFM technique, groove profile shapes, as it has been demonstrated in the present study. Today, using an appropriate AFM instrument and the respective method one has a possibility to determine with a superfine (atomic-scale) spatial resolution grating-like structures, i.e., their groove profiles including abrupt slopes and random nanoroughness. Moreover, such non-destructive AFM analysis is the only suitable one to apply to current production and evaluation of such complicated and expensive devices like as most of X-ray diffraction gratings are.
I thank David A. Content, John F. Seely, Tamara N. Berezovskaya, Vladislav A. Sharov for the information provided.
This work was partially supported by the Russian Foundation for Basic Research (RFBR) (Grant No. 20-02-00326) and the Russian Science Foundation (RSF) (Grant No. 19-12-00270) in the theoretical part.
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Her research interests include archaea metabolism, enzymes purification and characterization, gene regulation, carotenoids and bioplastics production, antioxidant\ncompounds, waste water treatments, and brines bioremediation.\nRosa María’s other roles include editorial board member for several journals related\nto biochemistry, reviewer for more than 60 journals (biochemistry, molecular biology, biotechnology, chemistry and microbiology) and president of several organizing committees in international meetings related to the N-cycle or respiratory processes.",institutionString:null,institution:{name:"University of Alicante",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null},{id:"15",title:"Chemical Biology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/15.jpg",isOpenForSubmission:!0,editor:{id:"441442",title:"Dr.",name:"Şükrü",middleName:null,surname:"Beydemir",slug:"sukru-beydemir",fullName:"Şükrü Beydemir",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00003GsUoIQAV/Profile_Picture_1634557147521",biography:"Dr. Şükrü Beydemir obtained a BSc in Chemistry in 1995 from Yüzüncü Yıl University, MSc in Biochemistry in 1998, and PhD in Biochemistry in 2002 from Atatürk University, Turkey. He performed post-doctoral studies at Max-Planck Institute, Germany, and University of Florence, Italy in addition to making several scientific visits abroad. He currently works as a Full Professor of Biochemistry in the Faculty of Pharmacy, Anadolu University, Turkey. Dr. Beydemir has published over a hundred scientific papers spanning protein biochemistry, enzymology and medicinal chemistry, reviews, book chapters and presented several conferences to scientists worldwide. He has received numerous publication awards from various international scientific councils. He serves in the Editorial Board of several international journals. 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He is a member of the Turkish Biochemical Society, American Chemical Society, and German Genetics society. Dr. Ekinci published around ninety scientific papers, reviews and book chapters, and presented several conferences to scientists. He has received numerous publication awards from several scientific councils. 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He worked on the structure-function relationships of glycoconjugates and his main project was the investigations on the biological roles of the de-N-glycosylation enzymes (Endo-N-acetyl-β-D-glucosaminidase and peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidase). From 2002 he contributes to the understanding of the Blood-brain barrier functioning using proteomics approaches. He has published more than 70 papers. 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Since then, he has been working as an Adjunct Professor in the same Department at the University of Pavia. His research activity during the first years was primarily focused on the purification and structural characterization of enzymes from animal and plant sources. During this period, Prof. Iadarola familiarized himself with the conventional techniques used in column chromatography, spectrophotometry, manual Edman degradation, and electrophoresis). Since 1995, he has been working on: i) the determination in biological fluids (serum, urine, bronchoalveolar lavage, sputum) of proteolytic activities involved in the degradation processes of connective tissue matrix, and ii) on the identification of biological markers of lung diseases. In this context, he has developed and validated new methodologies (e.g., Capillary Electrophoresis coupled to Laser-Induced Fluorescence, CE-LIF) whose application enabled him to determine both the amounts of biochemical markers (Desmosines) in urine/serum of patients affected by Chronic Obstructive Pulmonary Disease (COPD) and the activity of proteolytic enzymes (Human Neutrophil Elastase, Cathepsin G, Pseudomonas aeruginosa elastase) in sputa of these patients. More recently, Prof. Iadarola was involved in developing techniques such as two-dimensional electrophoresis coupled to liquid chromatography/mass spectrometry (2DE-LC/MS) for the proteomic analysis of biological fluids aimed at the identification of potential biomarkers of different lung diseases. He is the author of about 150 publications (According to Scopus: H-Index: 23; Total citations: 1568- According to WOS: H-Index: 20; Total Citations: 1296) of peer-reviewed international journals. 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She gained considerable experience in developing and validating new methodologies whose applications allowed her to determine both the amount of biomarkers (Desmosine and Isodesmosine) in the urine of patients affected by COPD, and the activity of proteolytic enzymes (HNE, Cathepsin G, Pseudomonas aeruginosa elastase) in the sputa of these patients. Simona Viglio was also involved in research dealing with the supplementation of amino acids in patients with brain injury and chronic heart failure. She is presently engaged in the development of 2-DE and LC-MS techniques for the study of proteomics in biological fluids. The aim of this research is the identification of potential biomarkers of lung diseases. 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He is especially interested in the genetic differentiation pattern and speciation process that correlate to the flashing pattern and mating behavior of some fireflies in Japan. He then worked for Olympus Corporation, a Japanese manufacturer of optics and imaging products, where he was involved in the development of luminescence technology and produced a bioluminescence microscope that is currently being used for gene expression analysis in chronobiology, neurobiology, and developmental biology. 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He is also a faculty member in the Molecular Oncology Program. He obtained his MSc and Ph.D. at Oregon State University and Texas Tech University, respectively. He pursued his postdoctoral studies at Rutgers University Medical School and the National Institutes of Health (NIH/NIDDK), USA. His research focuses on biochemistry, biophysics, genetics, molecular biology, and molecular medicine with specialization in the fields of drug design, protein structure-function, protein folding, prions, microRNA, pseudogenes, molecular cancer, epigenetics, metabolites, proteomics, genomics, protein expression, and characterization by spectroscopic and calorimetric methods.",institutionString:"University of Health Sciences",institution:null},{id:"180528",title:"Dr.",name:"Hiroyuki",middleName:null,surname:"Kagechika",slug:"hiroyuki-kagechika",fullName:"Hiroyuki Kagechika",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/180528/images/system/180528.jpg",biography:"Hiroyuki Kagechika received his bachelor’s degree and Ph.D. in Pharmaceutical Sciences from the University of Tokyo, Japan, where he served as an associate professor until 2004. He is currently a professor at the Institute of Biomaterials and Bioengineering (IBB), Tokyo Medical and Dental University (TMDU). From 2010 to 2012, he was the dean of the Graduate School of Biomedical Science. Since 2012, he has served as the vice dean of the Graduate School of Medical and Dental Sciences. He has been the director of the IBB since 2020. Dr. Kagechika’s major research interests are the medicinal chemistry of retinoids, vitamins D/K, and nuclear receptors. He has developed various compounds including a drug for acute promyelocytic leukemia.",institutionString:"Tokyo Medical and Dental University",institution:{name:"Tokyo Medical and Dental University",country:{name:"Japan"}}},{id:"40482",title:null,name:"Rizwan",middleName:null,surname:"Ahmad",slug:"rizwan-ahmad",fullName:"Rizwan Ahmad",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/40482/images/system/40482.jpeg",biography:"Dr. Rizwan Ahmad is a University Professor and Coordinator, Quality and Development, College of Medicine, Imam Abdulrahman bin Faisal University, Saudi Arabia. Previously, he was Associate Professor of Human Function, Oman Medical College, Oman, and SBS University, Dehradun. Dr. Ahmad completed his education at Aligarh Muslim University, Aligarh. He has published several articles in peer-reviewed journals, chapters, and edited books. His area of specialization is free radical biochemistry and autoimmune diseases.",institutionString:"Imam Abdulrahman Bin Faisal University",institution:{name:"Imam Abdulrahman Bin Faisal University",country:{name:"Saudi Arabia"}}},{id:"41865",title:"Prof.",name:"Farid A.",middleName:null,surname:"Badria",slug:"farid-a.-badria",fullName:"Farid A. Badria",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/41865/images/system/41865.jpg",biography:"Farid A. Badria, Ph.D., is the recipient of several awards, including The World Academy of Sciences (TWAS) Prize for Public Understanding of Science; the World Intellectual Property Organization (WIPO) Gold Medal for best invention; Outstanding Arab Scholar, Kuwait; and the Khwarizmi International Award, Iran. He has 250 publications, 12 books, 20 patents, and several marketed pharmaceutical products to his credit. He continues to lead research projects on developing new therapies for liver, skin disorders, and cancer. Dr. Badria was listed among the world’s top 2% of scientists in medicinal and biomolecular chemistry in 2019 and 2020. He is a member of the Arab Development Fund, Kuwait; International Cell Research Organization–United Nations Educational, Scientific and Cultural Organization (ICRO–UNESCO), Chile; and UNESCO Biotechnology France",institutionString:"Mansoura University",institution:{name:"Mansoura University",country:{name:"Egypt"}}},{id:"329385",title:"Dr.",name:"Rajesh K.",middleName:"Kumar",surname:"Singh",slug:"rajesh-k.-singh",fullName:"Rajesh K. Singh",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329385/images/system/329385.png",biography:"Dr. Singh received a BPharm (2003) and MPharm (2005) from Panjab University, Chandigarh, India, and a Ph.D. (2013) from Punjab Technical University (PTU), Jalandhar, India. He has more than sixteen years of teaching experience and has supervised numerous postgraduate and Ph.D. students. He has to his credit more than seventy papers in SCI- and SCOPUS-indexed journals, fifty-five conference proceedings, four books, six Best Paper Awards, and five projects from different government agencies. He is currently an editorial board member of eight international journals and a reviewer for more than fifty scientific journals. He received Top Reviewer and Excellent Peer Reviewer Awards from Publons in 2016 and 2017, respectively. He is also on the panel of The International Reviewer for reviewing research proposals for grants from the Royal Society. He also serves as a Publons Academy mentor and Bentham brand ambassador.",institutionString:"Punjab Technical University",institution:{name:"Punjab Technical University",country:{name:"India"}}},{id:"142388",title:"Dr.",name:"Thiago",middleName:"Gomes",surname:"Gomes Heck",slug:"thiago-gomes-heck",fullName:"Thiago Gomes Heck",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/142388/images/7259_n.jpg",biography:null,institutionString:null,institution:{name:"Universidade Regional do Noroeste do Estado do Rio Grande do Sul",country:{name:"Brazil"}}},{id:"336273",title:"Assistant Prof.",name:"Janja",middleName:null,surname:"Zupan",slug:"janja-zupan",fullName:"Janja Zupan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/336273/images/14853_n.jpeg",biography:"Janja Zupan graduated in 2005 at the Department of Clinical Biochemistry (superviser prof. dr. Janja Marc) in the field of genetics of osteoporosis. Since November 2009 she is working as a Teaching Assistant at the Faculty of Pharmacy, Department of Clinical Biochemistry. In 2011 she completed part of her research and PhD work at Institute of Genetics and Molecular Medicine, University of Edinburgh. She finished her PhD entitled The influence of the proinflammatory cytokines on the RANK/RANKL/OPG in bone tissue of osteoporotic and osteoarthritic patients in 2012. From 2014-2016 she worked at the Institute of Biomedical Sciences, University of Aberdeen as a postdoctoral research fellow on UK Arthritis research project where she gained knowledge in mesenchymal stem cells and regenerative medicine. She returned back to University of Ljubljana, Faculty of Pharmacy in 2016. She is currently leading project entitled Mesenchymal stem cells-the keepers of tissue endogenous regenerative capacity facing up to aging of the musculoskeletal system funded by Slovenian Research Agency.",institutionString:null,institution:{name:"University of Ljubljana",country:{name:"Slovenia"}}},{id:"357453",title:"Dr.",name:"Radheshyam",middleName:null,surname:"Maurya",slug:"radheshyam-maurya",fullName:"Radheshyam Maurya",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/357453/images/16535_n.jpg",biography:null,institutionString:null,institution:{name:"University of Hyderabad",country:{name:"India"}}},{id:"311457",title:"Dr.",name:"Júlia",middleName:null,surname:"Scherer Santos",slug:"julia-scherer-santos",fullName:"Júlia Scherer Santos",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/311457/images/system/311457.jpg",biography:"Dr. Júlia Scherer Santos works in the areas of cosmetology, nanotechnology, pharmaceutical technology, beauty, and aesthetics. Dr. Santos also has experience as a professor of graduate courses. Graduated in Pharmacy, specialization in Cosmetology and Cosmeceuticals applied to aesthetics, specialization in Aesthetic and Cosmetic Health, and a doctorate in Pharmaceutical Nanotechnology. Teaching experience in Pharmacy and Aesthetics and Cosmetics courses. She works mainly on the following subjects: nanotechnology, cosmetology, pharmaceutical technology, aesthetics.",institutionString:"Universidade Federal de Juiz de Fora",institution:{name:"Universidade Federal de Juiz de Fora",country:{name:"Brazil"}}},{id:"219081",title:"Dr.",name:"Abdulsamed",middleName:null,surname:"Kükürt",slug:"abdulsamed-kukurt",fullName:"Abdulsamed Kükürt",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRNVJQA4/Profile_Picture_2022-03-07T13:23:04.png",biography:"Dr. Kükürt graduated from Uludağ University in Turkey. He started his academic career as a Research Assistant in the Department of Biochemistry at Kafkas University. In 2019, he completed his Ph.D. program in the Department of Biochemistry at the Institute of Health Sciences. He is currently working at the Department of Biochemistry, Kafkas University. He has 27 published research articles in academic journals, 11 book chapters, and 37 papers. He took part in 10 academic projects. He served as a reviewer for many articles. He still serves as a member of the review board in many academic journals. His research interests include biochemistry, oxidative stress, reactive species, antioxidants, lipid peroxidation, inflammation, reproductive hormones, phenolic compounds, female infertility.",institutionString:"Kafkas University",institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"178366",title:"Associate Prof.",name:"Volkan",middleName:null,surname:"Gelen",slug:"volkan-gelen",fullName:"Volkan Gelen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/178366/images/system/178366.jpg",biography:"Volkan Gelen is a Physiology specialist who received his veterinary degree from Kafkas University in 2011. Between 2011-2015, he worked as an assistant at Atatürk University, Faculty of Veterinary Medicine, Department of Physiology. In 2016, he joined Kafkas University, Faculty of Veterinary Medicine, Department of Physiology as an assistant professor. Dr. Gelen has been engaged in various academic activities at Kafkas University since 2016. There he completed 5 projects and has 3 ongoing projects. He has 60 articles published in scientific journals and 20 poster presentations in scientific congresses. His research interests include physiology, endocrine system, cancer, diabetes, cardiovascular system diseases, and isolated organ bath system studies.",institutionString:"Kafkas University",institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"418963",title:"Dr.",name:"Augustine Ododo",middleName:"Augustine",surname:"Osagie",slug:"augustine-ododo-osagie",fullName:"Augustine Ododo Osagie",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/418963/images/16900_n.jpg",biography:"Born into the family of Osagie, a prince of the Benin Kingdom. I am currently an academic in the Department of Medical Biochemistry, University of Benin. Part of the duties are to teach undergraduate students and conduct academic research.",institutionString:null,institution:{name:"University of Benin",country:{name:"Nigeria"}}},{id:"192992",title:"Prof.",name:"Shagufta",middleName:null,surname:"Perveen",slug:"shagufta-perveen",fullName:"Shagufta Perveen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/192992/images/system/192992.png",biography:"Prof. Shagufta Perveen is a Distinguish Professor in the Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia. Dr. Perveen has acted as the principal investigator of major research projects funded by the research unit of King Saud University. She has more than ninety original research papers in peer-reviewed journals of international repute to her credit. She is a fellow member of the Royal Society of Chemistry UK and the American Chemical Society of the United States.",institutionString:"King Saud University",institution:{name:"King Saud University",country:{name:"Saudi Arabia"}}},{id:"49848",title:"Dr.",name:"Wen-Long",middleName:null,surname:"Hu",slug:"wen-long-hu",fullName:"Wen-Long Hu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/49848/images/system/49848.jpg",biography:"Wen-Long Hu is Chief of the Division of Acupuncture, Department of Chinese Medicine at Kaohsiung Chang Gung Memorial Hospital, as well as an adjunct associate professor at Fooyin University and Kaohsiung Medical University. Wen-Long is President of Taiwan Traditional Chinese Medicine Medical Association. He has 28 years of experience in clinical practice in laser acupuncture therapy and 34 years in acupuncture. He is an invited speaker for lectures and workshops in laser acupuncture at many symposiums held by medical associations. He owns the patent for herbal preparation and producing, and for the supercritical fluid-treated needle. Dr. Hu has published three books, 12 book chapters, and more than 30 papers in reputed journals, besides serving as an editorial board member of repute.",institutionString:"Kaohsiung Chang Gung Memorial Hospital",institution:{name:"Kaohsiung Chang Gung Memorial Hospital",country:{name:"Taiwan"}}},{id:"298472",title:"Prof.",name:"Andrey V.",middleName:null,surname:"Grechko",slug:"andrey-v.-grechko",fullName:"Andrey V. Grechko",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/298472/images/system/298472.png",biography:"Andrey Vyacheslavovich Grechko, Ph.D., Professor, is a Corresponding Member of the Russian Academy of Sciences. He graduated from the Semashko Moscow Medical Institute (Semashko National Research Institute of Public Health) with a degree in Medicine (1998), the Clinical Department of Dermatovenerology (2000), and received a second higher education in Psychology (2009). Professor A.V. Grechko held the position of Сhief Physician of the Central Clinical Hospital in Moscow. He worked as a professor at the faculty and was engaged in scientific research at the Medical University. Starting in 2013, he has been the initiator of the creation of the Federal Scientific and Clinical Center for Intensive Care and Rehabilitology, Moscow, Russian Federation, where he also serves as Director since 2015. He has many years of experience in research and teaching in various fields of medicine, is an author/co-author of more than 200 scientific publications, 13 patents, 15 medical books/chapters, including Chapter in Book «Metabolomics», IntechOpen, 2020 «Metabolomic Discovery of Microbiota Dysfunction as the Cause of Pathology».",institutionString:"Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology",institution:null},{id:"199461",title:"Prof.",name:"Natalia V.",middleName:null,surname:"Beloborodova",slug:"natalia-v.-beloborodova",fullName:"Natalia V. Beloborodova",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/199461/images/system/199461.jpg",biography:'Natalia Vladimirovna Beloborodova was educated at the Pirogov Russian National Research Medical University, with a degree in pediatrics in 1980, a Ph.D. in 1987, and a specialization in Clinical Microbiology from First Moscow State Medical University in 2004. She has been a Professor since 1996. Currently, she is the Head of the Laboratory of Metabolism, a division of the Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Moscow, Russian Federation. N.V. Beloborodova has many years of clinical experience in the field of intensive care and surgery. She studies infectious complications and sepsis. She initiated a series of interdisciplinary clinical and experimental studies based on the concept of integrating human metabolism and its microbiota. Her scientific achievements are widely known: she is the recipient of the Marie E. Coates Award \\"Best lecturer-scientist\\" Gustafsson Fund, Karolinska Institutes, Stockholm, Sweden, and the International Sepsis Forum Award, Pasteur Institute, Paris, France (2014), etc. Professor N.V. Beloborodova wrote 210 papers, five books, 10 chapters and has edited four books.',institutionString:"Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology",institution:null},{id:"354260",title:"Ph.D.",name:"Tércio Elyan",middleName:"Azevedo",surname:"Azevedo Martins",slug:"tercio-elyan-azevedo-martins",fullName:"Tércio Elyan Azevedo Martins",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/354260/images/16241_n.jpg",biography:"Graduated in Pharmacy from the Federal University of Ceará with the modality in Industrial Pharmacy, Specialist in Production and Control of Medicines from the University of São Paulo (USP), Master in Pharmaceuticals and Medicines from the University of São Paulo (USP) and Doctor of Science in the program of Pharmaceuticals and Medicines by the University of São Paulo. Professor at Universidade Paulista (UNIP) in the areas of chemistry, cosmetology and trichology. Assistant Coordinator of the Higher Course in Aesthetic and Cosmetic Technology at Universidade Paulista Campus Chácara Santo Antônio. Experience in the Pharmacy area, with emphasis on Pharmacotechnics, Pharmaceutical Technology, Research and Development of Cosmetics, acting mainly on topics such as cosmetology, antioxidant activity, aesthetics, photoprotection, cyclodextrin and thermal analysis.",institutionString:null,institution:{name:"University of Sao Paulo",country:{name:"Brazil"}}},{id:"334285",title:"Ph.D. Student",name:"Sameer",middleName:"Kumar",surname:"Jagirdar",slug:"sameer-jagirdar",fullName:"Sameer Jagirdar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/334285/images/14691_n.jpg",biography:"I\\'m a graduate student at the center for biosystems science and engineering at the Indian Institute of Science, Bangalore, India. I am interested in studying host-pathogen interactions at the biomaterial interface.",institutionString:null,institution:{name:"Indian Institute of Science Bangalore",country:{name:"India"}}},{id:"329795",title:"Dr.",name:"Mohd Aftab",middleName:"Aftab",surname:"Siddiqui",slug:"mohd-aftab-siddiqui",fullName:"Mohd Aftab Siddiqui",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329795/images/15648_n.jpg",biography:"Dr. Mohd Aftab Siddiqui is currently working as Assistant Professor in the Faculty of Pharmacy, Integral University, Lucknow for the last 6 years. He has completed his Doctor in Philosophy (Pharmacology) in 2020 from Integral University, Lucknow. He completed his Bachelor in Pharmacy in 2013 and Master in Pharmacy (Pharmacology) in 2015 from Integral University, Lucknow. He is the gold medalist in Bachelor and Master degree. He qualified GPAT -2013, GPAT -2014, and GPAT 2015. His area of research is Pharmacological screening of herbal drugs/ natural products in liver and cardiac diseases. He has guided many M. Pharm. research projects. He has many national and international publications.",institutionString:"Integral University",institution:null},{id:"255360",title:"Dr.",name:"Usama",middleName:null,surname:"Ahmad",slug:"usama-ahmad",fullName:"Usama Ahmad",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/255360/images/system/255360.png",biography:"Dr. Usama Ahmad holds a specialization in Pharmaceutics from Amity University, Lucknow, India. He received his Ph.D. degree from Integral University. Currently, he’s working as an Assistant Professor of Pharmaceutics in the Faculty of Pharmacy, Integral University. From 2013 to 2014 he worked on a research project funded by SERB-DST, Government of India. He has a rich publication record with more than 32 original articles published in reputed journals, 3 edited books, 5 book chapters, and a number of scientific articles published in ‘Ingredients South Asia Magazine’ and ‘QualPharma Magazine’. He is a member of the American Association for Cancer Research, International Association for the Study of Lung Cancer, and the British Society for Nanomedicine. Dr. Ahmad’s research focus is on the development of nanoformulations to facilitate the delivery of drugs that aim to provide practical solutions to current healthcare problems.",institutionString:"Integral University",institution:{name:"Integral University",country:{name:"India"}}},{id:"30568",title:"Prof.",name:"Madhu",middleName:null,surname:"Khullar",slug:"madhu-khullar",fullName:"Madhu Khullar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/30568/images/system/30568.jpg",biography:"Dr. Madhu Khullar is a Professor of Experimental Medicine and Biotechnology at the Post Graduate Institute of Medical Education and Research, Chandigarh, India. She completed her Post Doctorate in hypertension research at the Henry Ford Hospital, Detroit, USA in 1985. She is an editor and reviewer of several international journals, and a fellow and member of several cardiovascular research societies. Dr. Khullar has a keen research interest in genetics of hypertension, and is currently studying pharmacogenetics of hypertension.",institutionString:"Post Graduate Institute of Medical Education and Research",institution:{name:"Post Graduate Institute of Medical Education and Research",country:{name:"India"}}},{id:"223233",title:"Prof.",name:"Xianquan",middleName:null,surname:"Zhan",slug:"xianquan-zhan",fullName:"Xianquan Zhan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/223233/images/system/223233.png",biography:"Xianquan Zhan received his MD and Ph.D. in Preventive Medicine at West China University of Medical Sciences. He received his post-doctoral training in oncology and cancer proteomics at the Central South University, China, and the University of Tennessee Health Science Center (UTHSC), USA. He worked at UTHSC and the Cleveland Clinic in 2001–2012 and achieved the rank of associate professor at UTHSC. Currently, he is a full professor at Central South University and Shandong First Medical University, and an advisor to MS/PhD students and postdoctoral fellows. He is also a fellow of the Royal Society of Medicine and European Association for Predictive Preventive Personalized Medicine (EPMA), a national representative of EPMA, and a member of the American Society of Clinical Oncology (ASCO) and the American Association for the Advancement of Sciences (AAAS). He is also the editor in chief of International Journal of Chronic Diseases & Therapy, an associate editor of EPMA Journal, Frontiers in Endocrinology, and BMC Medical Genomics, and a guest editor of Mass Spectrometry Reviews, Frontiers in Endocrinology, EPMA Journal, and Oxidative Medicine and Cellular Longevity. He has published more than 148 articles, 28 book chapters, 6 books, and 2 US patents in the field of clinical proteomics and biomarkers.",institutionString:"Shandong First Medical University",institution:{name:"Affiliated Hospital of Shandong Academy of Medical Sciences",country:{name:"China"}}},{id:"297507",title:"Dr.",name:"Charles",middleName:"Elias",surname:"Assmann",slug:"charles-assmann",fullName:"Charles Assmann",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/297507/images/system/297507.jpg",biography:"Charles Elias Assmann is a biologist from Federal University of Santa Maria (UFSM, Brazil), who spent some time abroad at the Ludwig-Maximilians-Universität München (LMU, Germany). He has Masters Degree in Biochemistry (UFSM), and is currently a PhD student at Biochemistry at the Department of Biochemistry and Molecular Biology of the UFSM. His areas of expertise include: Biochemistry, Molecular Biology, Enzymology, Genetics and Toxicology. He is currently working on the following subjects: Aluminium toxicity, Neuroinflammation, Oxidative stress and Purinergic system. Since 2011 he has presented more than 80 abstracts in scientific proceedings of national and international meetings. Since 2014, he has published more than 20 peer reviewed papers (including 4 reviews, 3 in Portuguese) and 2 book chapters. He has also been a reviewer of international journals and ad hoc reviewer of scientific committees from Brazilian Universities.",institutionString:"Universidade Federal de Santa Maria",institution:{name:"Universidade Federal de Santa Maria",country:{name:"Brazil"}}},{id:"217850",title:"Dr.",name:"Margarete Dulce",middleName:null,surname:"Bagatini",slug:"margarete-dulce-bagatini",fullName:"Margarete Dulce Bagatini",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/217850/images/system/217850.jpeg",biography:"Dr. Margarete Dulce Bagatini is an associate professor at the Federal University of Fronteira Sul/Brazil. She has a degree in Pharmacy and a PhD in Biological Sciences: Toxicological Biochemistry. She is a member of the UFFS Research Advisory Committee\nand a member of the Biovitta Research Institute. She is currently:\nthe leader of the research group: Biological and Clinical Studies\nin Human Pathologies, professor of postgraduate program in\nBiochemistry at UFSC and postgraduate program in Science and Food Technology at\nUFFS. She has experience in the area of pharmacy and clinical analysis, acting mainly\non the following topics: oxidative stress, the purinergic system and human pathologies, being a reviewer of several international journals and books.",institutionString:"Universidade Federal da Fronteira Sul",institution:{name:"Universidade Federal da Fronteira Sul",country:{name:"Brazil"}}},{id:"226275",title:"Ph.D.",name:"Metin",middleName:null,surname:"Budak",slug:"metin-budak",fullName:"Metin Budak",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/226275/images/system/226275.jfif",biography:"Metin Budak, MSc, PhD is an Assistant Professor at Trakya University, Faculty of Medicine. He has been Head of the Molecular Research Lab at Prof. Mirko Tos Ear and Hearing Research Center since 2018. His specializations are biophysics, epigenetics, genetics, and methylation mechanisms. He has published around 25 peer-reviewed papers, 2 book chapters, and 28 abstracts. He is a member of the Clinical Research Ethics Committee and Quantification and Consideration Committee of Medicine Faculty. His research area is the role of methylation during gene transcription, chromatin packages DNA within the cell and DNA repair, replication, recombination, and gene transcription. His research focuses on how the cell overcomes chromatin structure and methylation to allow access to the underlying DNA and enable normal cellular function.",institutionString:"Trakya University",institution:{name:"Trakya University",country:{name:"Turkey"}}},{id:"243049",title:"Dr.",name:"Anca",middleName:null,surname:"Pantea Stoian",slug:"anca-pantea-stoian",fullName:"Anca Pantea Stoian",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/243049/images/system/243049.jpg",biography:"Anca Pantea Stoian is a specialist in diabetes, nutrition, and metabolic diseases as well as health food hygiene. She also has competency in general ultrasonography.\n\nShe is an associate professor in the Diabetes, Nutrition and Metabolic Diseases Department, Carol Davila University of Medicine and Pharmacy, Bucharest, Romania. She has been chief of the Hygiene Department, Faculty of Dentistry, at the same university since 2019. Her interests include micro and macrovascular complications in diabetes and new therapies. Her research activities focus on nutritional intervention in chronic pathology, as well as cardio-renal-metabolic risk assessment, and diabetes in cancer. She is currently engaged in developing new therapies and technological tools for screening, prevention, and patient education in diabetes. \n\nShe is a member of the European Association for the Study of Diabetes, Cardiometabolic Academy, CEDA, Romanian Society of Diabetes, Nutrition and Metabolic Diseases, Romanian Diabetes Federation, and Association for Renal Metabolic and Nutrition studies. She has authored or co-authored 160 papers in national and international peer-reviewed journals.",institutionString:null,institution:{name:"Carol Davila University of Medicine and Pharmacy",country:{name:"Romania"}}},{id:"279792",title:"Dr.",name:"João",middleName:null,surname:"Cotas",slug:"joao-cotas",fullName:"João Cotas",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/279792/images/system/279792.jpg",biography:"Graduate and master in Biology from the University of Coimbra.\n\nI am a research fellow at the Macroalgae Laboratory Unit, in the MARE-UC – Marine and Environmental Sciences Centre of the University of Coimbra. My principal function is the collection, extraction and purification of macroalgae compounds, chemical and bioactive characterization of the compounds and algae extracts and development of new methodologies in marine biotechnology area. \nI am associated in two projects: one consists on discovery of natural compounds for oncobiology. The other project is the about the natural compounds/products for agricultural area.\n\nPublications:\nCotas, J.; Figueirinha, A.; Pereira, L.; Batista, T. 2018. An analysis of the effects of salinity on Fucus ceranoides (Ochrophyta, Phaeophyceae), in the Mondego River (Portugal). Journal of Oceanology and Limnology. in press. DOI: 10.1007/s00343-019-8111-3",institutionString:"Faculty of Sciences and Technology of University of Coimbra",institution:null},{id:"279788",title:"Dr.",name:"Leonel",middleName:null,surname:"Pereira",slug:"leonel-pereira",fullName:"Leonel Pereira",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/279788/images/system/279788.jpg",biography:"Leonel Pereira has an undergraduate degree in Biology, a Ph.D. in Biology (specialty in Cell Biology), and a Habilitation degree in Biosciences (specialization in Biotechnology) from the Faculty of Science and Technology, University of Coimbra, Portugal, where he is currently a professor. In addition to teaching at this university, he is an integrated researcher at the Marine and Environmental Sciences Center (MARE), Portugal. His interests include marine biodiversity (algae), marine biotechnology (algae bioactive compounds), and marine ecology (environmental assessment). Since 2008, he has been the author and editor of the electronic publication MACOI – Portuguese Seaweeds Website (www.seaweeds.uc.pt). He is also a member of the editorial boards of several scientific journals. Dr. Pereira has edited or authored more than 20 books, 100 journal articles, and 45 book chapters. He has given more than 100 lectures and oral communications at various national and international scientific events. He is the coordinator of several national and international research projects. In 1998, he received the Francisco de Holanda Award (Honorable Mention) and, more recently, the Mar Rei D. Carlos award (18th edition). He is also a winner of the 2016 CHOICE Award for an outstanding academic title for his book Edible Seaweeds of the World. In 2020, Dr. Pereira received an Honorable Mention for the Impact of International Publications from the Web of Science",institutionString:"University of Coimbra",institution:{name:"University of Coimbra",country:{name:"Portugal"}}},{id:"61946",title:"Dr.",name:"Carol",middleName:null,surname:"Bernstein",slug:"carol-bernstein",fullName:"Carol Bernstein",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/61946/images/system/61946.jpg",biography:"Carol Bernstein received her PhD in Genetics from the University of California (Davis). She was a faculty member at the University of Arizona College of Medicine for 43 years, retiring in 2011. Her research interests focus on DNA damage and its underlying role in sex, aging and in the early steps of initiation and progression to cancer. In her research, she had used organisms including bacteriophage T4, Neurospora crassa, Schizosaccharomyces pombe and mice, as well as human cells and tissues. She authored or co-authored more than 140 scientific publications, including articles in major peer reviewed journals, book chapters, invited reviews and one book.",institutionString:"University of Arizona",institution:{name:"University of Arizona",country:{name:"United States of America"}}},{id:"182258",title:"Dr.",name:"Ademar",middleName:"Pereira",surname:"Serra",slug:"ademar-serra",fullName:"Ademar Serra",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/182258/images/system/182258.jpeg",biography:"Dr. Serra studied Agronomy on Universidade Federal de Mato Grosso do Sul (UFMS) (2005). He received master degree in Agronomy, Crop Science (Soil fertility and plant nutrition) (2007) by Universidade Federal da Grande Dourados (UFGD), and PhD in agronomy (Soil fertility and plant nutrition) (2011) from Universidade Federal da Grande Dourados / Escola Superior de Agricultura Luiz de Queiroz (UFGD/ESALQ-USP). Dr. Serra is currently working at Brazilian Agricultural Research Corporation (EMBRAPA). His research focus is on mineral nutrition of plants, crop science and soil science. Dr. Serra\\'s current projects are soil organic matter, soil phosphorus fractions, compositional nutrient diagnosis (CND) and isometric log ratio (ilr) transformation in compositional data analysis.",institutionString:"Brazilian Agricultural Research Corporation",institution:{name:"Brazilian Agricultural Research Corporation",country:{name:"Brazil"}}}]}},subseries:{item:{id:"9",type:"subseries",title:"Biotechnology - Biosensors, Biomaterials and Tissue Engineering",keywords:"Biotechnology, Biosensors, Biomaterials, Tissue Engineering",scope:"The Biotechnology - Biosensors, Biomaterials and Tissue Engineering topic within the Biomedical Engineering Series aims to rapidly publish contributions on all aspects of biotechnology, biosensors, biomaterial and tissue engineering. We encourage the submission of manuscripts that provide novel and mechanistic insights that report significant advances in the fields. Topics can include but are not limited to: Biotechnology such as biotechnological products and process engineering; Biotechnologically relevant enzymes and proteins; Bioenergy and biofuels; Applied genetics and molecular biotechnology; Genomics, transcriptomics, proteomics; Applied microbial and cell physiology; Environmental biotechnology; Methods and protocols. Moreover, topics in biosensor technology, like sensors that incorporate enzymes, antibodies, nucleic acids, whole cells, tissues and organelles, and other biological or biologically inspired components will be considered, and topics exploring transducers, including those based on electrochemical and optical piezoelectric, thermal, magnetic, and micromechanical elements. Chapters exploring biomaterial approaches such as polymer synthesis and characterization, drug and gene vector design, biocompatibility, immunology and toxicology, and self-assembly at the nanoscale, are welcome. Finally, the tissue engineering subcategory will support topics such as the fundamentals of stem cells and progenitor cells and their proliferation, differentiation, bioreactors for three-dimensional culture and studies of phenotypic changes, stem and progenitor cells, both short and long term, ex vivo and in vivo implantation both in preclinical models and also in clinical trials.",coverUrl:"https://cdn.intechopen.com/series_topics/covers/9.jpg",hasOnlineFirst:!1,hasPublishedBooks:!0,annualVolume:11405,editor:{id:"126286",title:"Dr.",name:"Luis",middleName:"Jesús",surname:"Villarreal-Gómez",slug:"luis-villarreal-gomez",fullName:"Luis Villarreal-Gómez",profilePictureURL:"https://mts.intechopen.com/storage/users/126286/images/system/126286.jpg",biography:"Dr. Luis Villarreal is a research professor from the Facultad de Ciencias de la Ingeniería y Tecnología, Universidad Autónoma de Baja California, Tijuana, Baja California, México. 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