Environmental Scanning Electron Microscopy in Cell Biology
Environmental scanning electron microscopy (ESEM) (1) is an imaging technique which allows hydrated, insulating samples to be imaged under an electron beam. The resolution afforded by this technique is higher than conventional optical microscopy but lower than conventional scanning electron microscopy (CSEM). The major advantage of the technique is the minimal sample preparation needed, making ESEM quick to use and the images less susceptible to the artifacts that the extensive sample preparation usually required for CSEM may introduce. Careful manipulation of both the humidity in the microscope chamber and the beam energy are nevertheless essential to prevent dehydration and beam damage artifacts. In some circumstances it is possible to image live cells in the ESEM (2).
In the following sections we introduce the fundamental principles of ESEM imaging before presenting imaging protocols for plant epidermis, mammalian cells, and bacteria. In the first two cases samples are imaged using the secondary electron (topographic) signal, whereas a transmission technique is employed to image bacteria.
Key wordsBacteria Hydrated STEM ESEM Plant epidermis Stomatal pores Mammalian cells Native state Surface imaging
The authors wish to thank T. W. Fairhead, D. J. Stokes, D. Waller, J. N. Skepper, A. Grant, Z. Wang, C. ffrench-Constant, A.A. R.Webb, P. J. Franks, and A. Luzhynskaya for their involvement with the development of these techniques.
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