Physics » "Scanning Electron Microscopy", book edited by Viacheslav Kazmiruk, ISBN 978-953-51-0092-8, Published: March 9, 2012 under CC BY 3.0 license

Chapter 3

In Situ Experiments in the Scanning Electron Microscope Chamber

By Renaud Podor, Johann Ravaux and Henri-Pierre Brau
DOI: 10.5772/36433

Article top

Overview

a) hot stage (FEI) b) Hot tension/compression stage integrated into an SEM (Kammrath & Weiss Co.) (After Biallas & Maier, 2007 ; Gorkaya et al., 2010).
Figure 1. a) hot stage (FEI) b) Hot tension/compression stage integrated into an SEM (Kammrath & Weiss Co.) (After Biallas & Maier, 2007 ; Gorkaya et al., 2010).
a) & (b) Single cell surgery without cell bursting using Si-Ti nanoneedle, (c) Force-cell deformation curve using Ti-Si and W2 nanoneedles at three different stages, i.e. (a) before penetration, (b) after penetration and (c) touching the substrate. (Ahmad et al., 2010).
Figure 2. a) & (b) Single cell surgery without cell bursting using Si-Ti nanoneedle, (c) Force-cell deformation curve using Ti-Si and W2 nanoneedles at three different stages, i.e. (a) before penetration, (b) after penetration and (c) touching the substrate. (Ahmad et al., 2010).
a) Simplified phase diagram for water indicating the ESEM domain (dot zone) and schemes to understand how isothermal or isobar experiments are performed. (b) Solubilisation and crystallization of NaCl directly observed in the ESEM chamber.
Figure 3. a) Simplified phase diagram for water indicating the ESEM domain (dot zone) and schemes to understand how isothermal or isobar experiments are performed. (b) Solubilisation and crystallization of NaCl directly observed in the ESEM chamber.

							In situ anther opening of C. angustifolia observed in LV-ESEM. 1) At the beginning, the valves of the anther are closed; 2) opening starts at the end of the stomium; 3) polyads are already seen; 4) opening proceeds till the valves are completely bent back and all eight polyads are presented (scale bar = 100µm). Time span from 1) to 3) was 25 min; 4) imaged 1 h after the start of the opening process (after Stabentheiner et al., 2010)
Figure 4. In situ anther opening of C. angustifolia observed in LV-ESEM. 1) At the beginning, the valves of the anther are closed; 2) opening starts at the end of the stomium; 3) polyads are already seen; 4) opening proceeds till the valves are completely bent back and all eight polyads are presented (scale bar = 100µm). Time span from 1) to 3) was 25 min; 4) imaged 1 h after the start of the opening process (after Stabentheiner et al., 2010)
Swelling kinetics of raw bentonite aggregates scale using ESEM-digital image analyses coupling (after Montes & Swelling, 2005).
Figure 5. Swelling kinetics of raw bentonite aggregates scale using ESEM-digital image analyses coupling (after Montes & Swelling, 2005).
ESEM micrographs of polyelectrolyte microcapsules suspended in double distilled water. Microcapsules were subjected to controlled dehydration in the ESEM sample chamber at T=5 °C. At an operating pressure of 800Pa, vesicles appeared as spherical structures. (a) Gradual decrease of the operating pressure to 350 Pa showed regular deformation of the microcaspsules (b to h)
Figure 6. ESEM micrographs of polyelectrolyte microcapsules suspended in double distilled water. Microcapsules were subjected to controlled dehydration in the ESEM sample chamber at T=5 °C. At an operating pressure of 800Pa, vesicles appeared as spherical structures. (a) Gradual decrease of the operating pressure to 350 Pa showed regular deformation of the microcaspsules (b to h)
Dehydration experiments performed on self-assembled organo-metallic compounds at T=22 °C and corresponding size modification versus water vapour pressure (Bonnefond, 2011).
Figure 7. Dehydration experiments performed on self-assembled organo-metallic compounds at T=22 °C and corresponding size modification versus water vapour pressure (Bonnefond, 2011).
Supersaturation versus temperature diagram for silver iodide (After Zimmermann et al., 2007).
Figure 8. Supersaturation versus temperature diagram for silver iodide (After Zimmermann et al., 2007).
Microdroplets growing and merging process under ESEM during increasing condensation by decreasing temperature. (After Jung & Bhushan, 2008)
Figure 9. Microdroplets growing and merging process under ESEM during increasing condensation by decreasing temperature. (After Jung & Bhushan, 2008)
Bright field image of 100 nm polystyrene latex spheres. Insert is the calibrated intensity corresponding to the dark line in the image (After Barkay (2010))
Figure 10. Bright field image of 100 nm polystyrene latex spheres. Insert is the calibrated intensity corresponding to the dark line in the image (After Barkay (2010))
~40 nm NaCl particles as the RH was increased past the deliquescence point. Water uptake [(a) →(b)] prior to full deliquescence (c) is clearly observed. (After Wise et al., 2008)
Figure 11. ~40 nm NaCl particles as the RH was increased past the deliquescence point. Water uptake [(a) →(b)] prior to full deliquescence (c) is clearly observed. (After Wise et al., 2008)
Growth of crystals in a borosilicate melt during 10 minutes isothermal heat treatment at 740 °C observed using the hot stage associated with the ESEM.
Figure 12. Growth of crystals in a borosilicate melt during 10 minutes isothermal heat treatment at 740 °C observed using the hot stage associated with the ESEM.
Decomposition of a uranium-cerium mixed oxalate observed during in situ heating in the ESEM chamber and relative size and shrinkage modifications.
Figure 13. Decomposition of a uranium-cerium mixed oxalate observed during in situ heating in the ESEM chamber and relative size and shrinkage modifications.
a) Sintering and grain growth of a uranium-cerium mixed oxide observed in situ in the ESEM chamber at T=1235 °C, after 55’, 70’, 90’, 95’, 130’, 140’ (a). Corresponding Relative (b) Shrinkage and Average grain diameter versus duration and (c) derived sintering map - Grain growth versus densification rate –
Figure 14. a) Sintering and grain growth of a uranium-cerium mixed oxide observed in situ in the ESEM chamber at T=1235 °C, after 55’, 70’, 90’, 95’, 130’, 140’ (a). Corresponding Relative (b) Shrinkage and Average grain diameter versus duration and (c) derived sintering map - Grain growth versus densification rate –

In Situ Experiments in theScanning Electron Microscope Chamber

Podor Renaud1, Ravaux Johann and Brau Henri-Pierre

1. Introduction

Since the first scanning electron microscope by Knoll (1935) and theoretical developments by von Ardenne (1938a, b) in the 30’s, this imaging technique has been widely used by generations of searchers from all the scientific domains to characterize the inner structure of matter. Even if the obtained information is essential for matter description or comprehension of matter transformation, the main constraints associated with classical electron microscopy, i.e. the necessity to work under vacuum and the necessity to prepare the sample before imaging, have always limited the possibilities to “post mortem” characterisation of samples and avoided observation of biological samples.

Electron microscopists early identified the necessity to undergo these limits. The development of a SEM chamber that is capable of maintaining a relatively high pressure and that allows imaging uncoated insulating samples began in the 70’s and has been “achieved” in the late 90’s – early 00’s (Stokes, 2008) with the commercialisation of the low-vacuum and environmental SEM. The availability of new generations of electron guns (and more particularly the field effect electron gun characterized by a very intense brightness), as well as the new generation of electronic columns that are now commonly associated with the environmental scanning electron microscopes opens new possibilities for material characterisation up to the nanometer scale. The development of this generation of microscopes have opened the door for performing real time experiments, using the electron microscope chamber as a microlab allowing direct observation of reactions at the micrometer scale. Many SEM providers or researchers have developed specific stages that can be used for the in situ experimentation in the scanning electron microscope chamber. This field is one of the most interesting uses of the ESEM that offers fantastic opportunities for matter properties characterisation. Even if numerous recent articles and reviews are dedicated to in situ experimentation in the VP/ESEM (Donald, 2003 ; Mendez-Vilas et al., 2009 ; Stokes, 2008 ; Stabentheiner et al., 2010 ; Gianola et al., 2011 ; Torres & Ramirez, 2011), no one describes all the possibilities of this technique. The present chapter will provide a large – and as exhaustive as possible – overview of the possibilities offered by the new SEM and ESEM generation in terms of “in situ experiments” focussing specifically on the more recent results (2000-2011).

This chapter will be split into five parts. We will first discuss the goals of in situ experimentation. Then, specific parts will be devoted to in situ mechanical tests, experiments under wet conditions, and a forth part dedicated to high temperature experiments in the SEM. Last, a specific part will be devoted to the “future” of in-SEM experiments. In each part, the main limits of the technique as well as the detection modes will be reported. Each part will be focussed on examples of the use of the technique for performing in situ experiments.

2. Goals and implementation requirements of in situ experimentation

The main goal of in situ experimentation in the SEM (or ESEM) chamber is to determine properties of matter through the study of its behaviour under constraint. This requires the combination of data collection over a given duration (on a unique sample) and image treatment for information extraction. The studied properties are generally related to microscopic phenomena and hardly assessable by other techniques. In situ experiment in the SEM chamber corresponds to both imaging systems in evolution under a constraint and imaging systems stabilized under controlled conditions.

To achieve this goal, several requirements are necessary:

  • The duration of the phenomenon to be observed must be suitable with the image recording time. If the system evolution is too fast, it will be impossible to record several images and observe this evolution. At the contrary, if the reaction kinetic is low, the time necessary for image recording will be too long and incompatible with experimentation. The high and low limits can be estimated ranging between 2 minutes and 48 hours.

  • The system must remain stable under the environmental conditions and/or irradiation by the electron beam during the time necessary for image recording. In the case of easily degradable samples, it is necessary to adjust the imaging conditions (high voltage, beam current, aperture, working distance, detector bias…) constantly, as the sample environmental conditions are modified during the experiment. Thus, the effect of the electron beam on the sample morphology modifications must be verified. Some authors report that it can act as an accelerator (Popma, 2002) or inhibitor (Courtois et al., 2011) of the observed reactions.

  • The image resolution must fit well with the size of details to be observed. Improvements in the image resolution have been achieved in the last decade thanks to the field effect emission guns. However, the presence of gas in the VP-SEM/ESEM chamber contributes to the incident electron beam scattering and subsequent degradation of the image resolution. Thus, the acquisition conditions must be adapted to the sample to be studied depending on the higher magnification to be reached.

  • The gaseous environmental conditions in which the studied system evolutes (or can be stabilized) must be reproduced in the SEM/LV-SEM/ESEM chamber. The development of the ESEM offers real new opportunities in term of composition of the atmosphere surrounding the sample. The large field detector and the gaseous secondary electron detector (Stokes, 2008) have been developed specifically for imaging under “high pressure” conditions (up to 300Pa and 3000Pa respectively) whatever the gas composition (air, water, He, He+H2 mixtures, O2). Other detectors have been developed for very specific applications (high temperature under vacuum (Nakamura et al., 2002), EBSD at high temperature (Fielden, 2005)).

  • The constraint in which the studied system evolutes (or can be stabilized) must also be reproduced in the microscope chamber. Some devices are commercialized by official sellers. Among them, we must report the Peltier stage for temperature control in the -10 to 60 °C range, hot stages for temperature control up to 1500 °C, stages for mechanical tests (Figure 1). Some authors have developed their own specific stages adapted to the problem to be treated (Fielden, 2005; Bogner et al., 2007). However, the development of miniaturized stages that can be positioned in the SEM chamber without creating perturbations on the incident electron beam can be really challenging. This will probably be a key in the development of in situ experimentation in the next years (Torres & Ramirez, 2011).

media/image2.png

Figure 1.

a) hot stage (FEI) b) Hot tension/compression stage integrated into an SEM (Kammrath & Weiss Co.) (After Biallas & Maier, 2007 ; Gorkaya et al., 2010).

The basis of in situ experimentation in the SEM is the study of the morphological modifications of the sample under constraint. Thus, this requires recording of numerous high quality images for image post treatment and data extraction in order to characterize the reaction or matter properties. The sample size can vary from 1µm to 50mm, and the image resolution is in the 1-10nm range, depending on recording conditions. The images are SEM images, i.e. with a large depth of field and with grey level contrasts. In-SEM experimentation can be extended to a wide range of applications, corresponding to very different materials (plants (Stabentheiner et al., 2010), food (Thiel et al., 2002 ; James, 2009), paper (Manero et al., 1998), soft matter, polymers, metals, ceramics, solids, liquids…) or problems (plant behaviour, chemical reactivity, properties characterization, sintering, grain growth, corrosion…). In the literature, the main part of the data reported has been acquired using an environmental scanning electron microscope.

3. In situ mechanical tests

Boehlert (2011) have recently underlined the interest of performing in situ mechanical tests in the SEM and summarized it as follows. “In situ scanning electron microscopy is now being routinely performed around the world to characterize the surface deformation behavior of a wide variety of materials. The types of loading conditions include simple tension, compression, bending, and creep as well as dynamic conditions including cyclic fatigue with dwell times. These experiments can be performed at ambient and elevated temperatures and in different environments and pressures. Most modern SEMs allow for the adaptation of heating and mechanical testing assemblies to the SEM stage, which allows for tilting and rotation to optimal imaging conditions as well as energy dispersive spectroscopy X-ray capture. Perhaps some of the most useful techniques involve acquisition of electron backscatter diffraction (EBSD) Kikuchi patterns for the identification of crystallographic orientations. Such information allows for the identification of phase transformations and plastic deformation as they relate to the local and global textures and other microstructural features. Understanding the microscale deformation mechanisms is useful for modeling and simulations used to link the microscale to the mesoscale behavior. In turn, simulations require verification through in situ microscale observations. Together simulations and in situ experimental verification studies are setting the stage for the future of material science, which undoubtedly involves accurate prediction of local and global mechanical properties and deformation behavior given only the processed microstructural condition”.

As a direct consequence of the great interest of the collected information, many different works from several scientific domains have been published for long. Thiel & Donald (1998) and Stabentheiner et al. (2010) describe the deformation of plants (carrots and leaves respectively) during room temperature tensile tests performed in the ESEM chamber. Similar tests are also reported with food (Stokes & Donald, 2000) and they are regularly performed on polymers (Poelt et al., 2010; Lin et al., 2010), composites (Schoßig et al., 2011) and metals (Boehlert et al., 2006; Gorkaya et al., 2010). Mechanical tests on metals, alloys and ceramics can also be performed at high temperature (Biallas & Maier, 2007; Chen & Boehlert, 2010). High temperature EDSB, developed by Seward et al. (2002), offers the possibility to observe phase transformations in materials as a function of temperature, as well as the direct visualization of the associated microstructural modifications (Seward et al., 2004).

media/image3.png

Figure 2.

a) & (b) Single cell surgery without cell bursting using Si-Ti nanoneedle, (c) Force-cell deformation curve using Ti-Si and W2 nanoneedles at three different stages, i.e. (a) before penetration, (b) after penetration and (c) touching the substrate. (Ahmad et al., 2010).

Several recently developed techniques allow characterizing materials at the nanometer scale through both technological miniaturization and advancements in imaging and small-scale mechanical testing. Ahmad et al. (2010) have developed a coupled ESEM-atomic force microscope to characterize single cells mechanical properties (Figure 2). This ESEM-nanomanipulation system allowed determining effects of internal influences (cell size and growth phases) and external influence (environmental conditions) on the cell strength. Gianola et al. (2011) reports the development of a quantitative in situ nanomechanical testing approach adapted to a dualbeam focused ion beam and scanning electron microscope. In situ tensile tests on 75 nm diameter Cu nanowhiskers as well as compression tests on nanoporous Au micropillars fabricated using FIB annular milling are reported, the scientific question being the mechanical behaviour of nanosize materials. Both examples probably represent what will be the future of in situ mechanical tests using scanning electron microscopes.

4. In situ experimentation under wet conditions

4.1. Conditions for experimentation

Combination of the use of the ESEM and a Peltier stage with the development of specific detectors allows the possibility to control both specimen temperature and water pressure around the sample (Leary & Brydson, 2010). Water can be condensed or evaporated on the demand from the sample (Figure 3). This allows performing in situ experiments in a temperature-pressure domain that is reported on Figure 3a (dot zone). An easy to perform experiment, illustrated by a 6 images series, corresponding to the NaCl dissolution (during the increasing of the water pressure in the ESEM chamber and consecutive water condensation, at constant temperature) in water followed by the crystallization of NaCl (decrease of the water pressure) is reported on Figure 3b. This example corresponds to an “isothermal experiment”. Another ways to work are to perform isobar experiments or to heat or cool a sample using a constant relative humidity (iso-RH experiments). These techniques allow the characterization of structural transitions of hydrated samples as a function of temperature (Bonnefond, 2011).

4.2. Biology and soft matter applications

This technique is particularly well adapted for the observation or experimentation on biological samples (Muscariello et al., 2005). Images of small and highly hydrated samples such as liposomes have been obtained by several authors (Perrie et al., 2007 ; Ruozi et al;, 2011) without any particular sample preparation. Perrie et al. (2007) have also been able to dynamically follow the hydration of lipid films and changes in liposome suspensions as water condenses onto, or evaporates from, the sample in real-time. The data obtained provides an insight into the resistance of liposomes to coalescence during dehydration, thereby providing an alternative assay for liposome formulation and stability (Perrie et al., 2010). However, Kirk et al. (2009) report that ESEM imaging of biological samples must remain combined with the classical techniques for sample preparation. Several works are specifically dedicated to in situ experimentation. Stabentheiner et al. (2010) state that “one unrivaled possibility of ESEM is the in situ investigation of dynamic processes that are impossible to access with CSEM where samples have to be fixed and processed”. These authors have studied the anther opening that is a highly dynamic process involving several tissue layers and controlled tissue desiccation. This phenomenon can be observed because the sample is very stable under the ESEM conditions (Figure 4). Another recent study is relative to the closure of stomatal pores by Mc Gregor & Donald (2010). Even if the possibility for experimentation on biological samples is clearly demonstrated, the authors outline the fact that the electron beam damages are important even at low accelerating voltage (Zheng et al., 2009). Another surprising example that can be reported is the direct observation of living acarids available online: in the movie, colonies of acarids are directly observed in the ESEM chamber under several conditions (FEI movie).

media/image4.png

Figure 3.

a) Simplified phase diagram for water indicating the ESEM domain (dot zone) and schemes to understand how isothermal or isobar experiments are performed. (b) Solubilisation and crystallization of NaCl directly observed in the ESEM chamber.

media/image5.png

Figure 4.

In situ anther opening of C. angustifolia observed in LV-ESEM. 1) At the beginning, the valves of the anther are closed; 2) opening starts at the end of the stomium; 3) polyads are already seen; 4) opening proceeds till the valves are completely bent back and all eight polyads are presented (scale bar = 100µm). Time span from 1) to 3) was 25 min; 4) imaged 1 h after the start of the opening process (after Stabentheiner et al., 2010)

4.3. Applications on cements

Several works have been performed in order to study the reactivity of cement materials versus humidity. Hydration or dehydration (Sorgi & De Gennaro, 2007; Fonseca & Jennings, 2010; Camacho-Bragado et al., 2011) of phases have been followed and used to extract kinetic parameters (Montes-Hernandez, 2002 ; Montes & Swelling, 2005 ; Maison et al., 2009), as reported on Figure 5. In this work, the author uses ESEM image series to determine a three-step mechanism for bentonite aggregates evolution with relative humidity corresponding to an arrangement of particles followed by a particle swelling and a full destructuration. In SEM experiments are also used to characterize chemical reactivity (Camacho-Bragado et al., 2011). It has been recently used to characterize reaction of fly ash activated by sodium silicate by Duchene et al. (2010). These authors have determined very accurately the different steps of the reaction determining that the sodium silicate activator dissolves rapidly and begins to bond fly ash particles. Open porosity was observed and it was rapidly filled with gel as soon as the liquid phase is able to reach the ash particle. The importance of the liquid phase is underlined as a fluid transport medium permitting the activator to reach and react with the fly ash particles. The reaction products had a gel like morphology and no crystallized phase was observed.

4.4. Hydration and dehydration experiments

As previously reported for liposomes, new opportunities for the study of polyelectrolyte microcapsules versus their resistance to relative humidity and temperature modifications are opened and under consideration. The image series reported on Figure 6 clearly illustrate the possibility to image the native soft capsule at high relative humidity without any deformation. When decreasing the water pressure near the capsule, the object is deformed and do not shrink as observed when it is heated in water at temperature higher than 25 °C (Basset et al., 2010). Thus, the walls of the object do not rearrange but collapse when submitted to a relative humidity decrease.

Similar tests have been performed on self-organized metal-organic framework compounds (Bonnefond, 2011). According to the image series reported on Figure 7, when the water pressure decreases, the size of sample remains constant up to a given water pressure (i.e. relative humidity) and for a transition pressure, the sample size decreases regularly. This can be associated to a local reorganisation in the sample that corresponds to a water loss associated to the sample collapsing The enthalpy of water ordering in the sample can be derived from the recorded image series as reported by Sievers et al.

media/image6.png

Figure 5.

Swelling kinetics of raw bentonite aggregates scale using ESEM-digital image analyses coupling (after Montes & Swelling, 2005).

media/image7.png

Figure 6.

ESEM micrographs of polyelectrolyte microcapsules suspended in double distilled water. Microcapsules were subjected to controlled dehydration in the ESEM sample chamber at T=5 °C. At an operating pressure of 800Pa, vesicles appeared as spherical structures. (a) Gradual decrease of the operating pressure to 350 Pa showed regular deformation of the microcaspsules (b to h)

media/image8.png

Figure 7.

Dehydration experiments performed on self-assembled organo-metallic compounds at T=22 °C and corresponding size modification versus water vapour pressure (Bonnefond, 2011).

The effect of dehydration on lamellar bones was also studied by in situ ESEM experiments (Utku et al., 2008). The obtained results indicate that dehydration affects the dimensions of lamellar bone in an anisotropic manner in longitudinal sections, whereas in transverse sections the extent of contraction is almost the same in both the radial and tangential directions.

An original work on the heterogeneous ice nucleation on synthetic silver iodide, natural kaolinite and montmorillonite particles has been performed using the “increasing water pressure at constant temperature” (Zimmermann et al., 2007) in the temperature range of 250–270 K. Ice formation was related to the chemical composition of the particles. The obtained data are in very good agreement with previous ones obtained by diffusion chamber measurements (Figure 8).

4.5. Characterization of surface wetting properties

Characterization of the wetting properties of surfaces through the formation of microdroplets or nanodroplets is another important investigation field that can be explored using the ESEM. A recent review by Mendez-Vilas et al. (2009) has highlighted the main fundamental and applied results. Several strategies for the contact angle between water and the surface determination are reported (Stelmashenko et al., 2001; Stokes, 2001; Lau et al., 2003; Wei, 2004; Yu et al., 2006; Jung & Bhushan, 2008; Rykaczewski & Scott, 2011). The investigation of the hydrophobicity and/or hydrophilicity of a catalyst layer have been performed using ESEM for the first time by Yu et al. (2006). These authors have determined the micro-contact angle distribution as a function of the catalyst microstructure. Microdroplets growing and merging process was observed directly in the ESEM chamber by Lau et al. (2003).

media/image9.png

Figure 8.

Supersaturation versus temperature diagram for silver iodide (After Zimmermann et al., 2007).

media/image10.png

Figure 9.

Microdroplets growing and merging process under ESEM during increasing condensation by decreasing temperature. (After Jung & Bhushan, 2008)

4.6. Using the Wet-STEM mode

The development of the Wet-STEM by Bogner et al. (2005, 2007) allows observing samples in the transmission mode in the ESEM chamber, and more particularly, it offers the possibility to image directly nanoparticles dispersed in a few micrometer thin water film (Bogner et al., 2008), emulsions or vesicles (Maraloiu et al., 2010), without removing the liquid surrounding the objects of interest. One must keep in mind that images with soft matter, and more generally sample sensitive to the electron beam are very hard to obtain. Nevertheless, this technique also opens new research fields using in situ experimentation that only begin to be explored for wettability or deliquescence studies. By combining Wet-STEM imaging with Monte-Carlo simulation (Figure 10), Barkay (2010) have studied the initial stages of water nanodroplet condensation over a nonhomogeneous holey thin film. This study has shown a preferred water droplet condensation over the residual water film areas in the holes and has provided corresponding droplet shape and contact angle. On a similar way, Wise et al. (2008) have studied water uptake by NaCl particles prior to deliquescence by varying the relative humidity in the Wet-STEM environment (Figure 11).

media/image11.png

Figure 10.

Bright field image of 100 nm polystyrene latex spheres. Insert is the calibrated intensity corresponding to the dark line in the image (After Barkay (2010))

media/image12.png

Figure 11.

~40 nm NaCl particles as the RH was increased past the deliquescence point. Water uptake [(a) →(b)] prior to full deliquescence (c) is clearly observed. (After Wise et al., 2008)

4.7. Development of specific materials for experimentation

Several specific devices have been developed to characterize specific properties or reactions. Two of them will be shortly described below.

Chen et al. (2011) have developed an experimental platform that can be used to investigate chemical reaction pathways, to monitor phase changes in electrodes or to investigate degradation effects in batteries. They have performed in situ experiment runs inside a scanning electron microscope (SEM) and tracked the morphology of an electrode including active and passive materials in real time. This work has been used to observe SnO2 during lithium uptake and release inside a working battery electrode.

Direct imaging of micro ink jets inside the ESEM chamber has been achieved using a specific device developed by Deponte et al. (2009), using a two-fluid stream consisting of a water inner core and a co-flowing outer gas sheath. ESEM images of water jets down to 700 nm diameter have been recorded. Details of the jet structure (the point of jet breakup, size and shape of the jet cone) can be measured. The authors conclude that ESEM imaging of liquid jets offers a valuable research tool for the study of aerosol production, combustion processes, ink-jet generation, and many other attributes of micro- and nanojet systems.

5. High temperature in the SEM

5.1. Application domains of HT-(E)SEM

Specific stages (and associated detectors) have been developed to heat samples up to 1500 °C directly in the microscope chamber (Knowles & Evans, 1997; Gregori et al., 2002). The environmental scanning electron microscope (ESEM) equipped with this heating stage is an excellent tool for the in situ and continuous observation of system modifications involved by temperature. It allows recording image series of the morphological changes of a sample during a heat treatment with both high magnification and high depth of focus. The experiments can be carried out to observe the influence of all these parameters on the studied phenomenon under various conditions (heating rates, atmosphere compositions, variable pressure, final temperature and heating time). Images have been recorded up to 1400 °C, with a decrease of the image resolution when the sample temperature increases (Podor et al., 2012). It is possible to work under vacuum (classical SEM) or under controlled atmosphere (H2O, O2, He+H2, N2, air...). Different types of studies have been reported, relative to corrosion of metals (Jonsson et al., 2011), oxidation of metals (Schmid et al., 2001a, 2001b ; Oquab & Monceau, 2001 ; Schmid et al., 2002 ; Abolhassani et al., 2003 ; Reichmann et al., 2008 ; Jonsson et al., 2009 ; Mège-Revil et al., 2009 ; Quémarda et al., 2009 ; Delehouzé et al., 2011), reactivity at high temperature (Maroni et al., 1999 ; Boucetta et al., 2010), phase changes (Fischer et al., 2004 ; Hung et al., 2007 ; Beattie & McGrady, 2009), hydrogen desorption (Beattie et al., 2009, 2011), redox reactions (Klemensø et al., 2006), microstructural modifications (Bestmann et al., 2005 ; Fielden, 2005 ; Yang, 2010), magnetic properties (Reichmann et al., 2011), sintering (Sample et al., 1996 ; Srinivasan, 2002 ; Marzagui & Cutard, 2004 ; Smith et al., 2006 ; Subramaniam, 2006 ; Courtois et al., 2011 ; Joly-Pottuz et al., 2011 ; Podor et al., 2012), thermal decomposition (Gualtieri et al., 2008 ; Claparède et al., 2011 ; Goodrich & Lattimer, 2011 ; Hingant et al., 2011), crystallisation (Gomez et al., 2009) in melts (Imaizumi et al., 2003 ; Hillers et al., 2007) and study of self-repairing – self-healing – properties of materials (Wilson & Case, 1997 ; Coillot et al., 2010a, 2010b, 2011) …

Even if numerous researchers are invested in HT-ESEM, only few of them have been successful in pursuing dynamic experiments at temperatures higher than 1100 °C. Two recent studies report experiments performed at T=1350 °C (Subramaniam, 2006) and 1450 °C (Gregori et al., 2002). However, the resolution of the images remains poor (more than 1µm) mainly due to water cooling induced vibrations. Furthermore, the precision on the measure of the sample temperature remains poor (temperature differences up to 150 °C with the expected temperature are sometimes measured). A recent device has been proposed by Podor et al. (2011) to overcome this difficulty.

A complete review specifically dedicated to in situ high temperature experimentation in the ESEM will be available soon. Several examples of in situ studies performed at high temperature in the ESEM chamber will be reported below, on the basis of original data acquired in our laboratory.

5.2. Investigation of the crystallization behaviour in silicate melts

The crystal growth and morphology during isothermal heating of glass melts can be directly observed using the hot stage associated with the ESEM. The image series reported on Figure 12 have been recorded during 10 minutes while heating the borosilicate melt sample isothermally at T=740 °C. The development of large crystals in the melt rapidly yields to the complete crystallization of the melt. The crystal morphology presents cells filled with a second phase and the crystal formation yields to the deformation of the sample surface. Hillers et al. (2007) have used such data to quantify the variation of crystal length with time. They have established that the growth is only linear during the first minutes; afterward the growth rate decreases progressively with time.

This technique can also be used to determine the temperature of formation of the first crystals at the melt surface and to observe their formation. In the case of glass-ceramics, the density of nuclei as well as their size and shape development can be directly observed and used for crystallization kinetic determination (Vigouroux et al., 2011, in prep).

media/image13.png

Figure 12.

Growth of crystals in a borosilicate melt during 10 minutes isothermal heat treatment at 740 °C observed using the hot stage associated with the ESEM.

5.3. Decomposition of compounds

In situ thermal decomposition of composites, oxalates, oxides have been reported by several authors. Images of the heat treatment of a mixed uranium-cerium oxalate grain from 25 °C to 1235 °C are gathered on Figure 13. Morphological changes with temperature are directly linked with the oxalate decomposition as stated by Hingant et al. (2011) in the temperature range 25-500 °C. The sample shrinkage observed when T>500 °C is probably related with the first stage of the sintering process – i.e. beginning of bond formation between the nanograins and with the oxide grain growth (that can not be directly observed at this stage by HT-ESEM, but that is confirmed by X-Ray diffraction). Such a process has also been recently reported by Claparede et al. (2011) and Joly-Pottuz et al. (2011).

media/image14.png

Figure 13.

Decomposition of a uranium-cerium mixed oxalate observed during in situ heating in the ESEM chamber and relative size and shrinkage modifications.

5.4. Study of sintering and grain growth

Several studies are relative to the sintering and grain growth processes in metals and ceramics. Depending on the system, the experiments have been performed in the temperature range 300-1450 °C. The main interest of these studies is the possibility of direct observation of the individual grain behaviour during heat treatment. The example that is reported on Figure 14a corresponds to the heat treatment of the grain decomposed in situ (Figure 13). The image resolution is high enough to observe the nanograins growth inside the square plate agglomerate. Consequently, relative shrinkage and average grain diameter are extracted by image processing (Figure 14b). Assuming that the final density of the agglomerate is 99%, the sintering map is directly derived from these experimental data (Figure 14c). Thus, in situ sintering experiments can allow the establishment of the trajectories of theoretical sintering. Such data have never been already reported in previous studies, mainly due to the poor resolution of the recorded images.

The effect of the electron beam on sintering is controversy. Indeed, Popma (2002) noted that a local sintering stop was achieved by focusing the electron beam at a certain position during the in situ sintering experiments in the ESEM (performed on ZrO2 nanolayers). On the contrary, Courtois et al (2011) performed experiments on the sintering of a lead phosphovanadate and concluded that the electric current induced by the electron beam was found to reduce the effective temperature of sintering by 50 to 150 °C as well as to accelerate the kinetics of shrinkage of a cluster composed of sub-micrometric grains of material. Such effects were not evidenced in our study: the local sintering on sample surface zones that were not observed (i.e. exposed to the electron beam) was identical to the local sintering determined on the observed zone.

media/image15.png

Figure 14.

a) Sintering and grain growth of a uranium-cerium mixed oxide observed in situ in the ESEM chamber at T=1235 °C, after 55’, 70’, 90’, 95’, 130’, 140’ (a). Corresponding Relative (b) Shrinkage and Average grain diameter versus duration and (c) derived sintering map - Grain growth versus densification rate –

6. Conclusions and perspectives

In situ scanning electron microscopy experimentation, that is generally associated with the use of the ESEM, allows the study of very different problems, the main limit being the availability of specific devices. Torres & Ramirez (2011) have written the best conclusion indicating that “the new generation of SEMs shows innovative hardware and software solutions that result in improved performance. This progress has turned the SEM into an extraordinary tool to develop more complex and realistic in situ experiments, achieving even at the subnanometer scale”. In the near future, new SEM imaging modes, nanomanipulation and nanofabrication technologies (Miller & Russell, 2007 ; Romano-Rodriguez & Hernandez-Ramirez, 2007 ; Wich et al., 2011) will make possible to replicate more closely the conditions as the ones associated to the problems to be treated. In situ ESEM will probably be used to overcome technical and fundamental challenges in many scientific domains. The recent developments of a high temperature stage in the FIB (Fielden, 2008), a new tomography mode in the ESEM (Jornsanoh et al., 2011) and of the atmospheric scanning electron microscope (Nishiyama et al, 2010 ; Suga et al, 2011) can be cited as examples for this future.

Acknowledgements

The authors want to thank all the co-workers of the studies cited in this chapter, and more particularly F. Bonnefond, H. Boucetta, C. Dejugnat, T. Demars, A. Monteiro and L. Claparède for providing the samples and challenging projects.

References

1 - S. Abolhassani, M. Dadras, M. Leboeuf, D. Gavillet, 2003 In situ study of the oxidation of Zircaloy-4 by ESEM. Journal of Nuclear Materials 321 70 77 .
2 - M. R. Ahmad, M. Nakajima, S. Kojima, M. Homma, T. Fukada, 2010 Single cell analysis inside ESEM- (ESEM)-nanomanipulator system, InTech, “Cutting Edge Nanotechnology” 978-9-53761-993-0 413 438 .
3 - Z. Barkay, 2010 Wettability study using transmitted electrons in environmental scanning electron microscope. Applied Physic Letters 96, 183109.
4 - C. Basset, C. Harder, C. Vidaud, C. Déjugnat, 2010 Design of Double Stimuli-Responsive Polyelectrolyte Microcontainers for Protein Soft Encapsulation. Biomacromolecules 11 806 814 .
5 - S. D. Beattie, G. S. Mc Grady, 2009 Hydrogen desorption studies of NaAlH4 and LiAlH4 by in situ heating in an ESEM. International Journal of Hydrogen Energy 34 9151 9156 .
6 - S. D. Beattie, H. W. Langmi, G. S. Mc Grady, 2009 In situ thermal desorption of H2 from LiNH2-2LiH monitored by environmental SEM. International Journal of Hydrogen Energy 34 376 379 .
7 - S. D. Beattie, U. Setthanan, G. S. Mc Grady, 2011 Thermal desorption of hydrogen from magnesium hydride (MgH2): An in situ microscopy study by environmental SEM and TEM. International Journal of Hydrogen Energy in press.
8 - M. Bestmann, S. Piazolo, C. J. Spiers, D. J. Prior, 2005 Microstructural evolution during initial stages of static recovery and recrystallization: new insights from in-situ heating experiments combined with electron backscatter diffraction analysis. Journal of Structural Geology 27 447 457 .
9 - G. Biallas, H. J. Maier, 2007 In-situ fatigue in an environmental scanning electron microscope- Potential and current limitations International Journal of Fatigue 29 1413 1425 .
10 - C. J. Boehlert, C. J. Cowen, S. Tamirisakandala, D. J. Mc Eldowney, D. B. Miracle, 2006 In situ scanning electron microscopy observations of tensile deformation in a boron-modified Ti-6Al-4V alloy. Scripta Materialia 55 465 468
11 - C. J. Boehlert, 2011 In situ scanning electron microscopy for understanding the deformation behaviour of structural materials. Seminarios Internacionales de Fronteras de la Ciencia de Materiales (http://www.youtube.com/watch?v=wH3EYxT_ysM)
12 - W. Chen, C. J. Boehlert, 2010 The 455 C tensile and fatigue behavior of boron-modified Ti-6Al-2Sn-4Zr-2Mo-0.1Si(wt.%). International Journal of Fatigue 32 799 807 .
13 - A. Bogner, A. Guimarães, R. C. O. Guimarães, A. M. Santos, G. Thollet, P. H. Jouneau, C. Gauthier, 2008 Grafting characterization of natural rubber latex particles: wet-STEM imaging contributions. Colloid and Polymer Science 286 1049 1059 .
14 - A. Bogner, P. H. Jouneau, G. Thollet, D. Basset, C. Gauthier, 2007 A history of scanning electron microscopy developments: Towards ‘‘wet-STEM’’ imaging. Micron 38 390 401 .
15 - A. Bogner, G. Thollet, D. Basset, P. H. Jouneau, C. Gauthier, 2005 Wet STEM: A new development in environmental SEM for imaging nano-objects included in a liquid phase Ultramicroscopy 104 290 301 .
16 - F. Bonnefond, 2011 Etude in situ de la déshydratation de composés organométalliques Master 1 thesis 30
17 - H. Boucetta, S. Schuller, J. Ravaux, R. Podor, 2010 Etude des mécanismes de formation des phases cristallines RuO2 dans les verres borosilicate de sodium. Proceeding of Matériaux
18 - G. A. Camacho-Bragado, F. Dixon, A. Colonna, 2011 Characterization of the response to moisture of talc and perlite in the environmental scanning electron microscope. Micron 42 257 262 .
19 - D. Chen, S. Indris, M. Schulz, B. Gamer, R. Mönig, 2011 In situ scanning electron microscopy on lithium-ion battery electrodes using an ionic liquid. Journal of Power Sources 196 6382 6387 .
20 - L. Claparède, N. Clavier, N. Dacheux, P. Moisy, R. Podor, J. Ravaux, 2011 Influence of crystallization state and microstructure on the chemical durability of cerium-neodynium mixed dioxides. Inorganic Chemistry, 50 18 9059 9072 .
21 - D. Coillot, R. Podor, F. O. Méar, L. Montagne, 2010 Characterisation of self-healing glassy composites by high-temperature environmental scanning electron microscopy (HT-ESEM). Journal of Electron Microscopy 59 359 366 .
22 - D. Coillot, F. O. Méar, R. Podor, L. Montagne, 2010 Autonomic Self-Repairing Glassy Materials. Advanced Functional Materials 20 24 4371 4374 .
23 - D. Coillot, F. O. Méar, R. Podor, L. Montagne, 2011 Influence of the Active Particles on the Self-Healing Efficiency in Glassy Matrix. Advanced Engineering Materials 13 426 435 .
24 - E. Courtois, G. Thollet, L. Campayo, S. Le Gallet, O. Bidault, F. Bernard, 2011 In situ study of the sintering of a lead phosphovanadate in an Environmental Scanning Electron Microscope. Solid State Ionics 186 53 58 .
25 - A. Delehouzé, F. Rebillat, P. Weisbecker, J. M. Leyssale, J. F. Epherre, C. Labrugère, G. L. Vignoles, 2011 Temperature induced transition from hexagonal to circular pits in graphite oxidation by O2. Applied Physics Letters 99 044102.
26 - D. P. De Ponte, R. B. Doak, M. Hunter, Z. Liu, U. Weierstall, J. C. H. Spence, 2009 SEM imaging of liquid jets. Micron 40 507 509 .
27 - A. M. Donald, 2003 The use of environmental scanning electron microscopy for imaging wet and insulating materials. Nature Materials 2 511 516 .
28 - J. Duchene, L. Duong, T. Bostrom, R. Frost, 2010 Microstructure study of early in situ reaction of fly ash geopolymer observed by ESEM. Waste Biomass Valorisation 1 367 377 .
30 - I. M. Fielden, 2005 Investigation of microstructural evolution by real time SEM of high temperature specimens. PhD thesis Sheffield Hallam University 170
31 - I. M. Fielden, 2008 In-Situ Focused Ion Beam (FIB) microscopy at high temperature. Electron Microscopy and Analysis Group.
32 - S. Fischer, K. Lemster, R. Kaegi, J. Kuebler, B. Grobety, 2004 In situ ESEM observation of melting silver and inconel on an Al2O3 powder bed. Journal of Electron Microscopy 53 393 396 .
33 - P. C. Fonseca, H. M. Jennings, 2010 The effect of drying on early-age morphology of C-S-H as observed in environmental SEM. Cement and Concrete Research 40 1673 1680 .
34 - D. S. Gianola, A. Sedlmayr, R. Mönig, C. A. Volkert, R. C. Major, E. Cyrankowski, S. A. S. Asif, O. L. Warren, O. Kraft, 2011 In situ nanomechanical testing in focused ion beam and scanning electron microscopes. Review of Scientific Instruments 82, 063901.
35 - L. S. Gómez, P. López-Arce, Buergo. M. Álvarez de, R. Fort, 2009 Calcium hydroxide nanoparticles crystallization on carbonates stone: dynamic experiments with heating/cooling and Peltier stage ESEM. Acta Microscopica 18 105 106 .
36 - T. W. Goodrich, B. Y. Lattimer, 2011 Fire Decomposition Effects on Sandwich Composite Materials. Composites A, doi:10.1016/j.compositesa.2011.03.007
37 - T. Gorkaya, T. Burlet, D. A. Molodov, G. Gottstein, 2010 Experimental method for true in situ measurements of shear-coupled grain boundary migration. Scripta Materialia 63 633 636 .
38 - G. Gregori, H. J. Kleebe, F. Siegelin, G. Ziegle, 2002 In situ SEM imaging at temperatures as high as 1450 C. Journal of Electron Microscopy 51 347 52 .
39 - A. F. Gualtieri, Gualtieri. M. Lassinantti, M. Tonelli, 2008 In situ ESEM study of the thermal decomposition of chrysotile asbestos in view of safe recycling of the transformation product. Journal of Hazardous Materials 156 260 266 .
40 - M. Hillers, G. Matzen, E. Veron, M. Dutreilh-Colas, A. Douy, 2007 Application of In Situ High-Temperature Techniques to Investigate the Effect of B2O3 on the Crystallization Behavior of Aluminosilicate E-Glass. Journal of the American Ceramic Society 90 720 726 .
41 - N. Hingant, N. Clavier, N. Dacheux, S. Hubert, N. Barré, R. Podor, L. Aranda, 2011 Preparation of morphology controlled Th1-xUxO2 sintered pellets from low-temperature precursors. Powder Technology 208 454 460 .
42 - J. H. H. Hung, Y. L. Chiu, T. Zhu, W. Gao, 2007 In situ ESEM study of partial melting and precipitation process of AZ91D. Asia-Pacific Journal of Chemical Engineering, Special Issue: Special issue for the Chemeca 2006 John A Brodie Medal Nominated Papers. 2 5 493 498
43 - K. Imaizumi, N. Matsuda, M. Otsuka, 2003 Coagulation/phase separation process in the silica/inorganic salt systems (1)-observation of state transformation Journal of Materials Science 38 2979 2986 .
44 - B. James, 2009 Advances in ‘‘wet’’ electron microscopy techniques and their application to the study of food structure Trends in Food Science & Technology 20 114 124 .
45 - L. Joly-Pottuz, A. Bogner, A. Lasalle, A. Malchere, G. Thollet, S. Deville, 2011 Improvements for imaging ceramics sintering in situ in ESEM. Journal of Microscopy, 244 93 100 .
46 - T. Jonsson, N. Folkeson, J. E. Svensson, L. G. Johansson, M. Halvarsson, 2011 An ESEM in situ investigation of initial stages of the KCl induced high temperature corrosion of a Fe-2.25Cr-1Mo steel at 400 C. Corrosion Science 53 2233 2246 .
47 - T. Jonsson, B. Pujilaksono, S. Hallström, J. Ågren, J. E. Svensson, L. G. Johansson, M. Halvarsson, 2009 An ESEM in situ investigation of the influence of H2O on iron oxidation at 500 C. Corrosion Science 51 1914 1924 .
48 - P. Jornsanoh, G. Thollet, J. Ferreira, K. Masenelli-Varlot, C. Gauthier, A. Bogner, 2011 Electron tomography combining ESEM and STEM: A new 3D imaging technique Ultramicroscopy, doi:10.1016/j.ultramic.2011.01.041
49 - Y. C. Jung, B. Bhushan, 2008 Wetting behaviour during evaporation and condensation of water microdroplets on superhydrophobic patterned surfaces Journal of Microscopy 229 127 140 .
50 - S. E. Kirk, J. N. Skepper, A. M. Donald, 2009 Application of environmental scanning electron microscopy to determine biological surface structure. Journal of Microscopy 233 205 224 .
51 - T. Klemensø, C. C. Appel, M. Mogensen, 2006 In Situ Observations of Microstructural Changes in SOFC Anodes during Redox Cycling. Electrochemical and Solid-State Letters 9 403 407
52 - M. Knoll, 1935 Aufladepototentiel une Sekündaremission elektronbestrahlter Körper. Zeitschrift fur technische Physik 16 467 475 .
53 - R. Knowles, B. Evans, 1997 High temperature specimen stage and detector for an ESEM. Patent WO 97/07526
54 - K. K. S. Lau, J. Bico, K. B. K. Teo, M. Chhowalla, G. A. J. Amaratunga, W. I. Milne, G. H. Mc Kinley, K. K. Gleason, 2003 Superhydrophobic Carbon Nanotube Forests. Nano Letters 3 1701 1705 .
55 - R. Leary, R. Brydson, 2010 Characterisation of ESEM conditions for specimen hydration control. Journal of Physics: Conference Series 241, 012024.
56 - T. Lin, D. Jia, M. Wang, 2010 In situ crack growth observation and fracture behavior of short carbon fiber reinforced geopolymer matrix composites. Materials Science and Engineering 527 2404 2407 .
57 - T. Maison, F. Laouafa, J. M. Fleureau, P. Delalain, 2009 Analyse aux échelles micro et macroscopique des mécanismes de dessiccation et de gonflement des sols argileux. Proceeding of the 19ème Congrès Français de Mécanique Marseille, 24 28
58 - J. M. Manero, D. V. Masson, M. Marsal, J. A. Planell, 1998 Application of the Technique of Environmental Scanning Electron Microscopy to the Paper Industry. ManeroJ. M.MassonD. V.MarsalM.PlanellJ. A. (1998). Application of the Technique of Environmental Scanning Electron Microscopy to the Paper Industry. Scanning . 21 36 39 .
59 - V. A. Maraloiu, M. Hamoudeh, H. Fessi, M. G. Blanchin, 2010 Study of magnetic nanovectors by Wet-STEM, a new ESEM mode in transmission. Journal of Colloid and Interface Science 352 386 392 .
60 - V. A. Maroni, M. Teplitsky, M. W. Rupich, 1999 An environmental scanning electron microscope study of the AgrBi-2223 composite conductor from 25 to 840 C. Physica C 313 169 174 .
61 - H. Marzagui, T. Cutard, 2004 Characterisation of microstructural evolutions in refractory castables by in situ high temperature ESEM. Journal of Materials Processing Technology 155-156, 1474 EOF -1481.
62 - J. E. Mc Gregor, A. M. Donald, 2010 ESEM imaging of dynamic biological processes: the closure of stomatal pores. Journal of Microscopy 239 135 141 .
63 - A. Mège-Revil, P. Steyer, G. Thollet, R. Chiriac, C. Sigala, J. C. Sanchéz-Lopéz, C. Esnouf, 2009 Thermogravimetric and in situ SEM characterisation of the oxidation phenomena of protective nanocomposite nitride films deposited on steel. Surface & Coatings Technology 204 893 901
64 - A. Mendez-Vilas, Jodar. Belen-Reyes, A. , M. L. Gonzalez-Martin, 2009 Ultrasmall Liquid Droplets on Solid Surfaces: Production, Imaging, and Relevance for Current Wetting Research. Mendez-VilasA.Belen-ReyesJodar.A.Gonzalez-MartinM. L. (2009). Ultrasmall Liquid Droplets on Solid Surfaces: Production, Imaging, and Relevance for Current Wetting Research. Small 5 12 1366 1390 .
65 - M. K. Miller, K. F. Russell, 2007 Atom probe specimen preparation with a dual beam SEM/FIB miller. Ultramicroscopy 107 761 766 .
66 - Montes-H, G. , 2005 Shrinkage measurements of bentonite using coupled environmental scanning electron microscopy and digital image analysis. Journal of Colloid and Interface Science 284 271 277 .
67 - G. Montes-Hernandez, 2002 Etude expérimentale de la sorption d’eau et du gonflement des argiles par microscopie électronique à balayage environnementale (ESEM) et l’analyse digitale d’images. Thèse de 3éme cycle 162
68 - L. Muscariello, F. Rosso, G. Marino, A. Giordano, M. Barbarisi, G. Cafiero, A. Barbarisi, 2005 A Critical Overview of ESEM Applications in the Biological Field. Journal of Cellular Physiology 205 328 334 .
69 - M. Nakamura, T. Isshiki, M. Tamai, K. Nishio, 2002 Development of a new heating stage equipped thermal electron filter for scanning electron microscopy. Proceeding of the 15th International Congress on Electron Microscopy Durban, South Africa.
70 - H. Nishiyama, M. Suga, T. Ogura, Y. Maruyama, M. Koizumi, K. Mio, S. Kitamura, C. Sato, 2010 Atmospheric scanning electron microscope observes cells and tissues in open medium through silicon nitride film. Journal of Structural Biology 169 438 449 .
71 - D. Oquab, D. Monceau, 2001 In-situ SEM study of cavity growth during high temperature oxidation of β-(Ni, Pd)Al. Scripta Materialia 44 2741 2746 .
72 - Y. Perrie, H. Ali, D. J. Kirby, A. U. R. Mohammed, S. E. Mc Neil, A. Vangala, 2010 Environmental Scanning Electron Microscope Imaging of Vesicle Systems in “Liposomes: Methods and Protocols, Volume 2: Biological Membrane Models” Methods in Molecular Biology 606 319 331.
73 - Y. Perrie, A. U. R. Mohammed, A. Vangala, S. E. Mc Neil, 2007 Environmental Scanning Electron Microscopy Offers Real-Time Morphological Analysis of Liposomes and Niosomes. Journal of Liposome Research 17 27 37 .
74 - R. Podor, N. Clavier, J. Ravaux, L. Claparéde, N. Dacheux, D. Bernache-Assollant, 2012 Dynamic aspects of cerium dioxide sintering: HT-ESEM study of grain growth and pore elimination. Journal of the European Ceramic Society, 32 353 362 .
75 - R. Podor, D. Pailhon, J. Ravaux, H. P. Brau, 2011 Porte-échantillon à thermocouple intégré. Demande de brevet français déposée le 21 juillet 2011 sous le n 11 56612.
76 - P. Poelt, A. Zankel, M. Gahleitner, E. Ingolic, C. Grein, 2010 Tensile tests in the environmental scanning electron microscope (ESEM)- Part I: Polypropylene homopolymers. Polymer 51 3203 3212 .
77 - R. L. W. Popma, 2002 Sintering characteristics of nano-ceramics, PhD thesis, University of Groningen.
78 - C. Proff, S. Abolhassani, M.M. Dadras, C. Lemaignan, 2010 In situ oxidation of zirconium binary alloys by environmental SEM and analysis by AFM, FIB, and TEM. Journal of Nuclear Materials 404 97 108.
79 - L. Quémarda, L. Desgranges, V. Bouineau, M. Pijolat, G. Baldinozzi, N. Millot, J. C. Nièpce, A. Poulesquen, 2009 On the origin of the sigmoid shape in the UO2 oxidation weight gain curves. Journal of the European Ceramic Society 29 2791 2798 .
80 - A. Reichmann, P. Poelt, C. Brandl, B. Chernev, P. Wilhelm, 2008 High-Temperature Corrosion of Steel in an ESEM With Subsequent Scale Characterisation by Raman Microscopy. Oxidation of Metals 78 257 266 .
81 - A. Reichmann, A. Zankel, H. Reingruber, P. Pölt, K. Reichmann, 2011 Direct observation of ferroelectric domain formation by environmental scanning electron microscopy. Journal of the European Ceramic Society, 31 15 2939 2942 .
82 - A. Romano-Rodriguez, F. Hernandez-Ramirez, 2007 Dual-beam focused ion beam (FIB): A prototyping tool for micro and nanofabrication. Microelectronic Engineering 84 789 792 .
83 - B. Ruozi, D. Belletti, A. Tombesi, G. Tosi, L. Bondioli, F. Forni, M. A. Vandelli, 2011 AFM, ESEM, TEM, and CLSM in liposomal characterization: a comparative study. International Journal of Nanomedicine 6 557 563 .
84 - K. Rykaczewski, J. H. J. Scott, 2011 Methodology for Imaging Nano-to-Microscale Water Condensation Dynamics on Complex Nanostructures. ACSNano 5 7 5962 5968 .
85 - D. R. Sample, P. W. Brown, J. P. Dougherty, 1996 Microstructural evolution of copper thick films observed by environmental scanning electron microscopy. Journal of the American Ceramic Society 79 1303 1306 .
86 - R. C. Schaller, N. Fukuta, 1979 Ice nucleation by aerosol particles: Experimental studies using a wedge-shaped ice thermal diffusion chamber. Journal of Atmospheric Sciences 36 1788 1802 .
87 - B. Schmid, Aas N. , Ø. Grong, Ø. Degard, R. , 2001 High-temperature oxidation of nickel and chromium studied with an in-situ environmental scanning electron microscope. Scanning 23 255 266 .
88 - B. Schmid, Aas N. , Ø. Grong, Ø. Degard, R. , 2001 In situ environmental scanning electron microscope observations of catalytic processes encountered in metal dusting corrosion on iron and nickel. Applied Catalysis 215 257 270 .
89 - B. Schmid, Aas N. , Ø. Grong, Ø. Degard, R. , 2002 High-Temperature Oxidation of Iron and the Decay of Wüstite Studied with in situ ESEM. Oxidation of Metals 57 115 130 .
90 - M. Schoßig, A. Zankel, C. Bieröge, P. Pölt, W. Grellmann, 2011 ESEM investigations for assessment of damage kinetics of short glass fibre reinforced thermoplastics- Results of in situ tensile tests coupled with acoustic emission analysis. Composites Science and Technology 71 257 265
91 - G. G. E. Seward, D. J. Prior, J. Wheeler, S. Celotto, D. J. M. Halliday, R. S. Paden, M. R. Tye, 2002 High-Temperature Electron Backscatter Diffraction and Scanning Electron Microscopy Imaging Techniques: In-situ Investigations of Dynamic Processes. Scanning 24 232 240 .
92 - G. G. E. Seward, S. Celotto, D. J. Prior, J. Wheeler, R. C. Pond, 2004 In situ SEM-EBSD observations of the hcp to bcc phase transformation in commercially pure titanium. Acta Materialia 52 821 832 .
93 - T. K. Sievers, F. Bonnefond, T. Demars, C. Genre, D. Meyer, R. Podor, Vapour pressure dependent size of coordination polymer network meso-particles. Advanced Materials (submitted)
94 - A. J. Smith, H. V. Atkinson, S. V. Hainsworth, A. C. F. Cocks, 2006 Use of a micromanipulator at high temperature in an environmental scanning electron microscope to apply force during the sintering of copper particles. Scripta Materialia 55 707 710 .
95 - C. Sorgi, V. De Gennaro, 2007 Analyse microstructurale au MEB environnemental d’une craie soumise à chargement hydrique et mécanique. Comptes-rendus Geosciences 339 468 481 .
96 - N. S. Srinivasan, 2002 Dynamic study of changes in structure and morphology during the heating and sintering of iron powder. Powder Technology 124 40 44 .
97 - E. Stabentheiner, A. Zankel, P. Pölt, 2010 Environmental scanning electron microscopy (ESEM)-a versatile tool in studying plants. Protoplasma 246 89 99 .
98 - N. A. Stelmashenko, J. P. Craven, A. M. Donald, E. M. Terentjev, B. L. Thiel, 2001 Topographic contrast of partially wetting water droplets in environmental scanning electron microscopy. Journal of Microscopy 204 172 183 .
99 - D. J. Stokes, A. M. Donald, 2000 In situ mechanical testing of dry and hydrated breadcrumb in the ESEM. Journal of Materials Science 35 599 607 .
100 - D. J. Stokes, 2001 Characterisation of Soft Condensed Matter and Delicate Materials Using Environmental Scanning Electron Microscopy (ESEM). Advanced Engineering Materials 3 126 130 .
101 - D. J. Stokes, 2008 Principles and practice of variable pressure/environmental scanning electron microscopy (VP/ESEM), John Wiley & Sons Ltd, The Atrium, Southern Gate, Chichester, West Sussex, UK.
102 - S. Subramaniam, 2006 In Situ High Temperature Environmental Scanning Electron Microscopic Investigations of Sintering Behavior in Barium Titanate. PhD thesis, University of Cincinnati, Cincinnati USA.
103 - M. Suga, H. Nishiyama, Y. Konyuba, S. Iwamatsu, Y. Watanabe, C. Yoshiura, T. Ueda, C. Sato, 2011 The Atmospheric Scanning Electron Microscope with open sample space observes dynamic phenomena in liquid or gas. Ultramicroscopy, doi:10.1016/j.ultramic.2011.08.001
104 - B. L. Thiel, A. M. Donald, 1998 In situ Mechanical Testing of Fully Hydrated Carrots (Daucus carota) in the Environmental SEM. Annals of Botany 82 727 733 .
105 - B. L. Thiel, D. J. Stokes, A. M. Donald, 2002 Application of Environmental Scanning Electron Microscopy to the Study of Food Systems. Microscopy and Microanalysis 8 960 961 .
106 - E. A. Torres, A. J. Ramirez, 2011 In situ scanning electron microscopy. Science and technology of welding and Joining 16 1 68 78 .
107 - F. S. Utku, E. Klein, H. Saybasili, C. A. Yucesoy, S. Weiner, 2008 Probing the role of water in lamellar bone by dehydration in the environmental scanning electron microscope. Journal of Structural Biology 162 361 367 .
108 - H. Vigouroux, E. Fargin, B. Le Garrec, M. Dussauze, V. Rodriguez, F. Adamietz, J. Ravaux, R. Podor, D. Vouagner, D. De Ligny, B. Champagnon, 2011 Phase Separation and Crystallization Mechanism in LiNbO -SiO2 Glasses. International conference on the chemistry of glasses 3
109 - H. Vigouroux, E. Fargin, B. Le Garrec, M. Dussauze, V. Rodriguez, F. Adamietz, J. Ravaux, R. Podor, D. Vouagner, D. De Ligny, B. Champagnon, 3In-Situ Study of LiNbO3 crystallization in lithium niobium Silicate glass ceramic. (in prep).
110 - Ardenne. M. von, 1938 Das Elektronen-Rastermikroskop. Praktische Ausführung. Zeitschrift fur technische Physik 19 407 416 .
111 - Ardenne. M. von, 1938 Das Elektronen-Rastermikroskop. Theoretische Grundlagen. Zeitschrift fur Physik 109 553 572 .
112 - Q. F. Wei, 2004 Surface characterization of plasma-treated polypropylene fibers. Materials Characterization 52 231 235 .
113 - T. Wich, C. Stolle, T. Luttermann, S. Fatikow, 2011 Assembly automation on the nanoscale. CIRP Journal of Manufacturing Science and Technology, doi:10.1016/j.cirpj.2011.03.003
114 - B. A. Wilson, D. E. Case, 1997 In situ microscopy of crack healing in borosilicate glass. Journal of Materials Science 32 3163 3175 .
115 - M. E. Wise, S. T. Martin, L. M. Russell, P. R. Buseck, 2008 Water uptake by NaCl particles prior to deliquescence and the phase rule. Aerosol Science and Technology 42 4 281 294 .
116 - J. Yang, 2010 In-situ High Resolution SEM Imaging with Heating Stage. Scanning Electron Microscopes (SEM) from Carl Zeiss.
117 - H. M. Yu, C. Ziegler, M. Oszcipok, M. Zobel, C. Hebling, 2006 Hydrophilicity and hydrophobicity study of catalyst layers in proton exchange membrane fuel cells. Electrochimica Acta 51 1199 1207 .
118 - T. Zheng, K. W. Waldron, A. M. Donald, 2009 Investigation of viability of plant tissue in the environmental scanning electron microscopy. Planta 230 1105 1113 .
119 - F. Zimmermann, M. Ebert, A. Worringen, L. Schutz, S. Weinbruch, 2007 Environmental scanning electron microscopy (ESEM) as a new technique to determine the ice nucleation capability of individual atmospheric aerosol particles. Atmospheric Environment 41 8219 8227 .
120 - Reactivity of a salt with silicate melt at high temperature http://www.dailymotion.com/icsmweb#videoId=xjknrt
121 - Sintering of CeO2 at T=1200 C http://www.youtube.com/watch? 4ijIUdQe3M4
122 - Self-healing of a metal-glass composite at high temperature http://www.dailymotion.com/icsmweb#videoId=xjknpp
123 - Deformation of vesicles during dehydration http://www.dailymotion.com/icsmweb#videoId=xjk75u
124 - NaCl solubility and precipitation in water http://www.dailymotion.com/icsmweb#videoId=xk22i9