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Abstract
Electron Microscopy by John Bozzola and Lonnie Russell continues to be a text of choice for instruction in classes that cover beginning electron microscopy for biologists and should also be on the bookshelf of all technologists that work with biological specimens for transmission or scanning EM. The first chapter in the book presents a brief, but interesting, history of the development of instrumentation and techniques for specimen preparation and sets the stage for the wealth of practical information that is presented in the remaining twenty chapters. These chapters are laid out in a logical chronological fashion of exactly what one must do to start with an unfixed specimen and end with a publication quality micrograph.
... TEM has resolution of up to 0.1 nm, which is thousand times greater than light microscope. This is possible because of the short wavelength of electron beam (2). With TEM, significant advancements in analyses of cellular compartments, such as cytoskeleton, membranes, organelles, cilia, etc. have been made (3). ...
... The electron beam must be able to penetrate the specimen. For this reason, biological specimen must be very thin, not thicker than 100 nm (2). Although TEM is more than appropriate device to study the internal structure of biological cells, conditions inside TEM and preparation of specimens may damage the cells. ...
... The goal of fixation is to preserve biological specimens from damage, to maintain its natural conditions, and to stabilize molecules against disruption by subsequent procedures such as dehydration and resin infiltration (4). There are two variations of this procedure: chemical fixation and cryo-fixation (2). For maximum tissue preservation it is important to choose proper method of fixation corresponding to the type of samples and structures of interest that need to be visualized. ...
Background purpose: The revolution in microscopy came in 1930 with the invention of electron microscope. Since then, we can study specimens on ultrastructural and even atomic level. Besides transmission electron microscopy (TEM), for which specimen preparation techniques will be described in this article, there are also other types of electron microscopes that are not discussed in this review. Materials and methods: Here, we have described basic procedures for TEM sample preparation, which include tissue sample preparation, chemical fixation of tissue with fixatives, cryo-fixation performed by quick freezing, dehydration with ethanol, infiltration with transitional solvents, resin embedding and polymerization, processing of embedded specimens, sectioning of samples with ultramicrotome, positive and negative contrasting of samples, immunolabeling, and imaging. Conclusion: Such collection of methods can be useful for novices in transmission electron microscopy.
Electron microscopy (EM), in its various flavors, has significantly contributed to our understanding of lipid droplets (LD) as central organelles in cellular metabolism. For example, EM has illuminated that LDs, in contrast to all other cellular organelles, are uniquely enclosed by a single phospholipid monolayer, revealed the architecture of LD contact sites with different organelles, and provided near‐atomic resolution maps of key enzymes that regulate neutral lipid biosynthesis and LD biogenesis. In this review, we first provide a brief history of pivotal findings in LD biology unveiled through the lens of an electron microscope. We describe the main EM techniques used in the context of LD research and discuss their current capabilities and limitations, thereby providing a foundation for utilizing suitable EM methodology to address LD‐related questions with sufficient level of structural preservation, detail, and resolution. Finally, we highlight examples where EM has recently been and is expected to be instrumental in expanding the frontiers of LD biology.
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