By Geoffrey Bourne (Eds.)
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Extra info for Cytology and Cell Physiology
A number of reasons might be cited to rationalize these discord ant results including: (a) the method of cell disruption, (b) processing of abnormally large amounts of tissue for the isolation of mitochondria, and (c) adequacy of the fixation, embedding, and sectioning procedures used prior to electron microscopic examination. Regardless of which of these processes is operative, the fact remains that mitochondria have been 2. CENTRIFUGAL ISOLATION OF SUBCELLULAR COMPONENTS 35 isolated from liver homogenized in isotonic sucrose in a good state of preservation and high degree of purity as shown in Fig.
20 WALTER C. SCHNEIDER AND EDWARD L. KUFF part responsible for the lack of consideration of isolation methods. They argued, on the basis of their observations of living cells under the micro scope, that disruption of the cell wall produced immediate and irrevers ible changes in the subcellular components. Consequently, isolation methods were dismissed a priori as studies of artifacts. One cytologist, however, Robert Russell Bensley, compared the cell to a watch and maintained that one could not understand the work ings of either unless the composition and function of each component part was established.
It should be noted that if nuclei are isolated in sucrose solution, their appearance is very similar to that seen in the living cell. When such nuclei are disrupted, however, over 6 0 % of the nucleoprotein is nonsedimentable at 60,000 g whereas in nuclei prepared in saline over 9 0 % of the nucleoprotein is sedimented at low speeds (5000 g or less) (Schneider and Hogeboom, 1951). It is clear that the chromosomal material is in a considerably different form under these two conditions. It would seem desirable to reinvestigate the isolation of chromosomes using synchronous cultures of cells since large numbers of cells in a specific stage of mitosis could be obtained.
Cytology and Cell Physiology by Geoffrey Bourne (Eds.)