The rat liver cell nuclear imprint as a standard for DNA measurements

Standards are used in DNA cytophotometry to determine the diploid reference value. Fixation and staining protocols have to be standardized because numerous sources of variation influence staining intensity of DNA. When Feulgen stained imprints of rat liver cell nuclei are used as an external DNA standard a significant difference of reference value of up to 60% between imprints, from the same liver and with all sources of variation minimized, is demonstrated. Different possible sources of intra-imprint variation were examined but its exact nature remains unknown. Due to this inter-imprint variation, the DNA-reference value from a single liver imprint is only an approximation with unknown error of the true diploid reference value. By drawing an aselective random sample of N diploid cells in M imprints the sampling distribution of TIOD (mean Total Integrated Optical Density) will have its expectation identical to the diploid reference value and is approached by TIOD (mean TIOD) with the confidence interval defined by the number, M, of imprints measured. The use of TIOD as an estimate of diploid reference value with known precision is proposed as a first approach for the definition of a standard for DNA measurements.