The quality of the image is affected not only by fixation of the material, but also by the conditions of its study in the scanning electron microscope. If the sprayed layer of carbon or heavy metal is thin there is the risk of charging the specimen during rapid scanning with the electron beam, and this interferes with the quality of the image. The study of biological objects is best carried out at two nominal values of the accelerating voltage (for example, 10 and 4 kV), for in this way errors during interpretation of the results can be avoided. Neither the thickness of the layer of carbon or heavy metal or different nominal values of the accelerating voltage can affect the reliability of the information obtained if in each concrete case the physical basis of image formation and the experimental conditions are allowed for in the analysis. Museum material fixed in ordinary formalin can be used for study in the scanning electron microscope.
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