Direct counting and sizing of mitochondria in solution.
Resistive particle counting has been developed for the accurate sizing and counting of mitochondria in solution. The normal detection limit with a 30 micro aperture is 0.48 micro diameter, or 0.056 micro(3) particle volume The mean volume of rat liver mitochondria was 0.42 micro(3) or 0.93 micro in diameter. The average value for numbers of particles per milligram of mitochondrial protein was 4.3 x 10(3), and per gram of rat liver was about 11 x 10(10). These values compare satisfactorily with those derived by light microscopy and electron microscopy. The mean volume for mitochondria from rat heart was 0 60 micro(3) and from rat kidney cortex, 0.23 micro(3). These values agree within 15% of those determined by electron microscopy of whole tissue. Mitochondrial fragility and contaminating subcellular organelles were shown to have little influence on the experimentally determined size distributions The technique may be applied to rapid swelling studies, as well as to estimations of the number and size of mitochondria from animals under different conditions such as liver regeneration and hormonal, pathological, or drug-induced states Mitochondrial DNA, RNA, cytochrome c-oxidase, cytochrome (a / a(3)), and iron were nearly constant per particle over large differences in particle size. Such data may be particularly valuable for biogenesis studies and support the hypothesis that the net amount per particle of certain mitochondrial constituents remains constant during mitochondrial growth and enlargement
D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D002374:Catalase / Q000032:analysis; D003585:Cell Biology / Q000295:instrumentation*; D002499:Centrifugation, Density Gradient; D003580:Cytochromes / Q000032:analysis; D004247:DNA / Q000032:analysis; D003576:Electron Transport Complex IV / Q000032:analysis; D005952:Glucose-6-Phosphatase / Q000032:analysis; D066298:In Vitro Techniques; D007668:Kidney / Q000166:cytology; D008297:Male; D008928:Mitochondria; D008930:Mitochondria, Liver; D008931:Mitochondria, Muscle; D008933:Mitochondrial Swelling; D009206:Myocardium / Q000166:cytology; D012313:RNA / Q000032:analysis; D051381:Rats; D012996:Solutions; D013053:Spectrophotometry
D003580:Cytochromes; D012996:Solutions; D012313:RNA; D004247:DNA; D002374:Catalase; D003576:Electron Transport Complex IV; D000135:Acid Phosphatase; D005952:Glucose-6-Phosphatase