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In Situ Study of Live Specimens in an Environmental Scanning Electron Microscope

Published online by Cambridge University Press:  02 May 2013

Eva Tihlaříková ,
Vilém Neděla  and
Makoto Shiojiri
Eva Tihlaříková
Affiliation:
Institute of Scientific Instruments of the ASCR, v.v.i., Královopolská 147, Brno 612 64, Czech Republic
Vilém Neděla*
Affiliation:
Institute of Scientific Instruments of the ASCR, v.v.i., Královopolská 147, Brno 612 64, Czech Republic
Makoto Shiojiri
Affiliation:
Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan
*
*Corresponding author.[email protected]
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Abstract

In this paper we introduce new methodology for the observation of living biological samples in an environmental scanning electron microscope (ESEM). The methodology is based on an unconventional initiation procedure for ESEM chamber pumping, free from purge–flood cycles, and on the ability to control thermodynamic processes close to the sample. The gradual and gentle change of the working environment from air to water vapor enables the study of not only living samples in dynamic in situ experiments and their manifestation of life (sample walking) but also its experimentally stimulated physiological reactions. Moreover, Monte Carlo simulations of primary electron beam energy losses in a water layer on the sample surface were studied; consequently, the influence of the water thickness on radiation, temperature, or chemical damage of the sample was considered.

Keywords

live biological samplehigh-pressure environmenthydration systemin situ observationMonte Carlo simulationmethodology
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 19 , Issue 4 , August 2013 , pp. 914 - 918
Copyright
Copyright © Microscopy Society of America 2013 

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Footnotes

†

These authors contributed equally to this work.

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