Studies on cell structure by the freezing‐drying method VI. The preparation and properties of mitochondria

Abstract

As indicated iii the preceding paper, the mitochoiidria of the liver of the guinea pig and rabbit are wholly insoluble in a 0.85 solution of sodium chloride. In order to obtain mitochondria for examination, it is only necessary to extract by diff erential centrifugation the emulsion of guinea pig liver described in the previous article.The emulsion is prepared by grinding or crushing in a mortar fresh liver which has been it-ashed free of blood by perfusion, or by kneading through bolting silk. This emulsion, as stated in a previous paper, contains fragments of small aggregations of liver cells, individual liver cells in great abundance, large numbers of nuclei released from cytoplasm, a few blood corpuscles and much reticulum arid mitochondria. The salt solution used for this preparation has a p H of about 6, so has a slightly acid reaction. Distinctly alkaline Ringer solntions are quite unsuited for the purpose and salt solutions made with conductivity wafer arc not so successful as solutions made with ordinary distilled water, having the reaction indicated above.Centrifugation of this cmulsion at low speed for a period of 3 niinntes sedimenls the majority of cell masses. If an attempt be made to get out the suspended cells at one operation, a great loss of mitochondria ensues, so it is better to rid the suspension of the groups of intact cells, individual '

Bioanalysis of Eukaryotic Organelles

“…Hoerr investigated the total composition of “fatty substances” in enriched mitochondria fraction from guinea pig and rabbit liver mitochondria in 1934. 141 The authors found 43.6% of fatty substances, primarily glycerides, i n the enriched m itochondria and investigated the presence of lecithin and cephalin. C laude in vestigated the phospholipid composition of subcellular f ractions of rat lymphoid cells in 1944 and found the phospholipid composition of lipids from mitochondria to be 75–80% using elemental analysis.…”

A history of research on yeasts 4: cytology part II, 1950–1990

“…But ‘it was Bensley's45 pioneering work’ in the early 1930s, Lehninger tells us, ‘that presaged the confluence of cytological work on mitochondria with biochemical research on respiration’ (120 p. 4). Bensley, appreciating the potential value for biochemical studies of obtaining large quantities of isolated mitochondria, tried to isolate them from liver cells by differential centrifugation25 and Claude46 succeeded in doing so in 194644. At the Rockefeller Institute in New York, cytologists, such as Claude, combined with biochemists, such as Hogeboom47 101, 102, and this association soon led to the finding that mitochondria are the principal sites of respiration111.…”

A short history of tissue fractionation.

“…Other landmarks in the history of tissue fractionation are the attempts made in the laboratory of R . R. Bensley to separate and to analyze pure mitochondria (8), and the isolation of microsomes by Claude (9) . It is interesting that the latter discovery concerned a cell component that could not be seen as such in the light microscope and was, therefore, not known to exist (except in those cells where it forms large basophilic masses, or ergastoplasm) .…”