Abstract
In this article we review the application and procedures involved in scanning electron microscope (SEM) to observe biological and live tissues through using SEM at high resolution. We discuss practical methods for optimizing tissue preservation to achieve the two principal goals of biological specimen preparation: (a) preserving biological structures as close to their living configuration as possible, and (b) rendering them visible with the desired imaging method. We also review and discuss the relative merits of different fixing (chemical fixation and cryofixation), drying (air-drying, critical point-drying, freeze-drying and chemical-drying) and coating procedures of biological specimens with metals to facilitate visualization in the SEM.
Highlights
Today, the Scanning Electron Microscope is utilized in materials, chemical and physic sciences, and in diverse fields such as medical sciences and biology
One of the most challenges in interpreting scanning electron microscope (SEM) images of biological specimens is to be able to distinguish between those features that reflect the native structure and those that are artificially created during processing
The problem of distinguishing real structures from artifacts is compounded by the fact that most microscopic methods from images not of biological structures directly, but stains, dyes, or coatings that are added to the specimen to render the parts of interest visible, but which themselves are artifacts [4]
Summary
The Scanning Electron Microscope (hereinafter abbreviated to SEM) is utilized in materials, chemical and physic sciences, and in diverse fields such as medical sciences and biology. Biological Samples and Live Tissues for SEM closed to 99% of chemical composition Z=120 and include the major elements (H, C, N, O) and minor elements (Na, Mg, Si, P, S., Cl, K, and Ca). Some types of biological tissues or specimens will require less stringent processing to preserve their structure. The first and most important step after sample selection for the study of living tissues is their rapid and suitable fixation method. The text by Glauert and Lewis [13] gives in-depth attention to all aspects of fixation and embedment of biological samples In this part, the techniques will be discussed primarily in regard to preparing specimens for SEM as well.
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