Fixation for Electron Microscopy by M. A. Hayat PDF

By M. A. Hayat

ISBN-10: 0123339200

ISBN-13: 9780123339201

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Alternatively, osmolarity can be adjusted by adding electrolytes such as NaCl and CaCl 2. These substances are added to the buffer before adjusting the pH, since they may change the final pH. Care is required in selecting the type and concentration of the substance to be added. If there is any indication that ionizing substances will have an adverse effect, nonelectrolytes should be used. In some 34 2. Factors Affecting the Quality of Fixation cases, monovalent ions are preferred over divalent ions, for the latter are more 2+ 2 + likely to produce ion effects.

214), and concentrated HCl (36-38%). Sodium cacodylate contains arsenic and thus is a health hazard. If inhaled or absorbed through skin, it can cause dermatitis, liver and kidney inflammation, etc. Hands should be protected by gloves and fume hoods should be used for weighing out the reagent and preparing the buffer solution. The reagent should not come in contact with acids in order to avoid the production of arsenic gas (also see Weakley, 1977). Since cacodylate reacts with H 2S , the two should not be used in the same solution.

This decreased leakage of proteins allows more cross-linking to occur. It has been shown that the addition of CaCl 2 to the fixative solution suppressed extraction of proteins in erythrocytes (Tooze, 1964). As stated earlier, divalent ions are assumed to be effective in reducing the protein repulsive force in the membrane system and provide some cross-linking between the separate, negatively charged protein chains. This cross-linking probably causes membrane stabilization, for proteins are anchored 2+ more firmly.

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Fixation for Electron Microscopy by M. A. Hayat

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