Simultaneous Measurement of DAPI-Sulforhodamine 101 Stained Nuclear DNA and Protein in Higher Plants by Flow Cytometry
作者:
UlrichWolfgang,
期刊:
Biotechnic&Histochemistry
(Taylor Available online 1992)
卷期:
Volume 67,
issue 2
页码: 73-78
ISSN:1052-0295
年代: 1992
DOI:10.3109/10520299209110011
出版商: Taylor&Francis
关键词: DAPI-sulforhodamine staining;nuclear DNA;nuclear protein;plant DNA and protein;flow cytometry
数据来源: Taylor
摘要:
A 2-step staining procedure is presented for simultaneous measurement of nuclear DNA and protein content in higher plants by flow cytometry. To release nuclei, plant tissues were chopped and stirred in the presence of the DNA specific fluorochrome 4′, 6-diamidino-2-phenylindole (DAPI) and the nonionic detergent Triton X-100. Plant protoplasts were stirred in the DAPI dye solution with the detergent. After a short incubation period a second dye solution containing DAPI and the protein fluorochrome sulforhodamine 101 (SR 101) without detergent was added. Following another incubation, and after filtration through nylon gauze, the highly fluorescent nuclei were analyzed with an impulse cytophotometer. Accurate bivariate DNA-protein histograms were obtained with CV-values of about 2% or less for the 2C-peak of the univariate DNA parameter. The method presented here can be used for basic and applied cytogenetic studies of higher plants, for characterization of subcompartments of the cell cycle phases, or for examination of heterogeneity in plant tissues.
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