Journal
MICROSCOPY AND MICROANALYSIS
Volume 19, Issue 4, Pages 914-918Publisher
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S1431927613000603
Keywords
live biological sample; high-pressure environment; hydration system; in situ observation; Monte Carlo simulation; methodology
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Funding
- Grant Agency of the Czech Republic [GAP 102/10/1410]
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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.
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