Absolute DNA values from Feulgen microspectrophotometric measurements and quantitative electron microscopy. A comparison of two species.
Quantitative electron microscopy (QEM) and microspectrophotometry were used to correlate the Feulgen stain absorption values to the calculated picograms of DNA. Measurements were made in human lymphocytes, rainbow trout lymphocytes and nuclei of trout erythrocytes. The median dry weight of the nucleus, as determined by QEM, was 35.9 pg for a human lymphocyte and 30.5 pg for a trout lymphocyte. Using Salzman's value of 20% DNA per chromosome (i.e., chromatin), a human lymphocyte nucleus thus contains 7.18 pg of DNA and a trout lymphocyte nucleus 6.1 pg of DNA. The mean Feulgen absorption value of the nucleus, given in arbitrary units (AU), was 14.5 for a human lymphocyte, 12.7 for a trout lymphocyte and 12.0 for a trout erythrocyte. From these values, it was derived that each picogram of DNA of a human lymphocyte nucleus is represented by 2.02 arbitrary Feulgen units while the values for trout nuclei were 2.08 AU and 1.97 AU. On the average, we find that each picogram of DNA is represented by two arbitrary Feulgen units in our microspectrophotometric measurements.
D000818:Animals; D004396:Coloring Agents; D004247:DNA / Q000032:analysis*; D004912:Erythrocytes / Q000032:analysis; D006801:Humans; D008214:Lymphocytes / Q000032:analysis; D008854:Microscopy, Electron; D012394:Rosaniline Dyes; D013045:Species Specificity; D013053:Spectrophotometry; D014337:Trout
D004396:Coloring Agents; C007722:Feulgen stain; D012394:Rosaniline Dyes; D004247:DNA