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Application Note
Biological samples span an incredible range of size scales, composition, structures, and morphology. Over the last few decades two complementary microscopy techniques for biological specimens have moved to the forefront, confocal laser scanning microscopy and scanning electron microscopy (SEM). With respect to the SEM many biological specimens share a similar set of issues, namely low conductivity of the sample and the typical glass substrates.
Introduction
Generally samples examined in the SEM need to be electrically conducting in order to minimize charge buildup on the sample from the electron beam. Charge buildup can severely degrade the resultant image data. [1] Advances in SEM to address a wider range of samples have led to brighter sources, ield emission ilaments, low vacuum also termed environmental SEM (eSEM), and low voltage SEM (LV-SEM). Three approaches can be employed to minimize charging. One approach is to metal coat the sample with an inert metal like gold. Another option is to increase the pressure in the sample chamber (eSEM) so that the gas molecules balance the charge. The third option is to decrease the electron beam voltage (LV-SEM) so that the beam energy is at the charge equilibrium point.
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