Biological Specimen Preparation for Transmission Electron Microscopy

Abstract

An introduction to fixation and embedding procedures I and their safe use in the laboratory 1.1 The scope of this book I 1.2 Criteria for the good preservation of ukrastructure 4 1.3 Artefacts in electron micrographs 6 1.4 Safety precautions in the electron microscope laboratory 9 1.4.1 General safety precautions 9 1.4.2 Hazards from electrical equipment and fire 10 1.4.3 Safe procedures for handling animals II 1.4.4 Dangers from sharp implements 12 1.4.5 Safe procedures with chemicals 12 1.4.5a Labelling for toxidty and flammabilivy 12 1.4.5b The safe handling of chemicals 15 1.4.5c Storage and disposal of chemicals 16 1.4.6 Hazards with liquid nitrogen II 1.4.7 First-aid in the laboratory 18 2 Fixatives 2 f 2. / An introduction to fixation 21 2.1.1 The development of fixatives for electron microscopy 22 2.1.2 The chemistry and physics of fixation 24 2.1.3 The penetration of fixatives 25 2.1.4 The choice of aldehyde fixative 27 2.2 The safe preparation and disposal of fixatives 29 2.2.1 Preparation of a fixative 30 2.2.2 Disposal of fixatives 32 2.2.3 Spillages 33 xiv Contents 2.3 Vehicles for fixatives 2.3.1 The choice of buffer 2.3.2 Osmolarity " 35 2.3.3 The preparation of buffers 37 2.4 Glutaraldehyde fixatives 2.4.1 The effects of glutaraldehyde on cells and tissues 40 2.4.1 a Reactions of glutaraldehyde with proteins 40 2.4.1b Reactions of glutaraldehyde with other cell constituents 44 2.4.1c The effects of the osmolarity of glutaraldehyde fixatives 44 2.4.2 The preparation of glutaraldehyde fixatives 45

Suppressors of the Chloroplast Protein Import Mutant tic40 Reveal a Genetic Link between Protein Import and Thylakoid Biogenesis

“…The grids were viewed under a JEOL JEM-1400 electron microscope at 80 kV. Digital images were recorded using a SIS digital camera and iTEM software (Glauert and Lewis, 1998;Hyman and Jarvis, 2011).…”

Light and electron microscopic study on the effect of antischizophrenic drugs on the structure of seminiferous tubules of adult male albino rats

“…Ultrathin sections (70-90 nm) were cut using an ultramicrotome. Uranyl acetate and lead citrate were used for staining of ultrathin sections [19] and studied by means of JEOL 1010 electron microscope (JEOL, Tokyo, Japan) at the Mycology and Regional Biotechnology Center, Al Azhar University, Cairo, Egypt.…”

Protective role of coenzyme Q 10 against high cholesterol dietinduced histological and biochemical changes in cerebellar cortex of adult albino rats

“…Then, they were postfixed in 1% osmium tetroxide in the same buffer at 4°C, dehydrated, and embedded in epoxy resin [17]. Semithin sections (1μm thick) were stained with 1% toluidine blue for light microscopic examination [18].…”