Get 20M+ Full-Text Papers For Less Than $1.50/day. Start a 14-Day Trial for You or Your Team.

Learn More →

Standardization of high‐resolution flow cytometric DNA analysis by the simultaneous use of chicken and trout red blood cells as internal reference standards

Standardization of high‐resolution flow cytometric DNA analysis by the simultaneous use of... Determination of nuclear DNA content by flow cytometry requires comparison with a reference standard. The use of external standards such as lymphocytes or granulocytes is time‐consuming and inaccurate. Chicken red blood cells (CRBC) have a DNA content of 35% of the human diploid value and have been widely used as internal standard. The ratio calculated on the basis of the peak channel numbers of the standard and the sample and used to indicate the DNA content (DNA ratio) is, however, very sensitive to changes in the zero level adjustment of the flow cytometer. If two internal standards are used the DNA ratio becomes independent of the zero level. Rainbow trout red blood cells (TRBC) have a DNA content of 80% of human diploid cells. A mixture of CRBC and TRBC was prepared and stored in small aliquots at —80°C. This mixture was added to the sample before staining. The day‐to‐day variation of the DNA ratio obtained by use of the two standards was smaller than that obtained by CRBC alone. The possibility of sex related differences in DNA content of CRBC and TRBC was examined. The results indicated that a new batch of standards should be tested against the old batch to avoid the introduction of a systematic error. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Cytometry Part A Wiley

Standardization of high‐resolution flow cytometric DNA analysis by the simultaneous use of chicken and trout red blood cells as internal reference standards

Loading next page...
 
/lp/wiley/standardization-of-high-resolution-flow-cytometric-dna-analysis-by-the-tUSWEptn4r

References (25)

Publisher
Wiley
Copyright
Copyright © 1983 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1552-4922
eISSN
1552-4930
DOI
10.1002/cyto.990030504
pmid
6839881
Publisher site
See Article on Publisher Site

Abstract

Determination of nuclear DNA content by flow cytometry requires comparison with a reference standard. The use of external standards such as lymphocytes or granulocytes is time‐consuming and inaccurate. Chicken red blood cells (CRBC) have a DNA content of 35% of the human diploid value and have been widely used as internal standard. The ratio calculated on the basis of the peak channel numbers of the standard and the sample and used to indicate the DNA content (DNA ratio) is, however, very sensitive to changes in the zero level adjustment of the flow cytometer. If two internal standards are used the DNA ratio becomes independent of the zero level. Rainbow trout red blood cells (TRBC) have a DNA content of 80% of human diploid cells. A mixture of CRBC and TRBC was prepared and stored in small aliquots at —80°C. This mixture was added to the sample before staining. The day‐to‐day variation of the DNA ratio obtained by use of the two standards was smaller than that obtained by CRBC alone. The possibility of sex related differences in DNA content of CRBC and TRBC was examined. The results indicated that a new batch of standards should be tested against the old batch to avoid the introduction of a systematic error.

Journal

Cytometry Part AWiley

Published: Mar 1, 1983

Keywords: ; ; ; ;

There are no references for this article.