PMID- 6185286
OWN - NLM
STAT- MEDLINE
DCOM- 19830324
LR  - 20071114
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 3
IP  - 4
DP  - 1983 Jan
TI  - RNA and DNA content of isolated cell nuclei measured by multiparameter flow 
      cytometry.
PG  - 269-75
AB  - Nuclei were isolated from various cell types including Chinese hamster ovary 
      (CHO) and L1210 leukemia cell lines, primary cultures of fibroblasts, 
      nonstimulated and stimulated human lymphocytes and mouse liver cells, by using 
      different isolation techniques. The isolated nuclei were subsequently stained 
      with acridine orange (AO) and their fluorescence was measured by flow cytometry. 
      Various procedures designed to stain DNA versus RNA differentially with AO were 
      tested, and the staining of isolated nuclei was compared with that of whole 
      cells. Control incubations with RNase and DNase were performed to estimate in 
      whole cells and in nuclei the contribution of DNA and RNA to the fluorescence 
      intensity at the respective wavelength bands of maximum emission for DNA (F530) 
      and RNA (F greater than 600). Depending on the cell type, 10-20% of total cell 
      RNase-sensitive F greater than 600 is localized in the nuclei. The 
      RNase-resistant portion of F greater than 600 of isolated nuclei represents the 
      stainability of DNA. Suppression of cell proliferation in subconfluent cultures 
      results in a decrease in both whole cell and in nuclear RNA content. 
      Nonstimulated lymphocyte nuclei have considerably lower RNA content than nuclei 
      from lymphocytes stimulated by pokeweed mitogen. Two subpopulations of nuclei 
      having the same (2C) DNA content but differing in RNA content, are present in 
      mouse liver; the cells entering S phase originate from the high RNA population.
FAU - Piwnicka, M
AU  - Piwnicka M
FAU - Darzynkiewicz, Z
AU  - Darzynkiewicz Z
FAU - Melamed, M R
AU  - Melamed MR
LA  - eng
GR  - CA-14134/CA/NCI NIH HHS/United States
GR  - CA-23296/CA/NCI NIH HHS/United States
GR  - CA-28704/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
RN  - 63231-63-0 (RNA)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Animals
MH  - Cell Nucleus/*analysis
MH  - Cells, Cultured
MH  - Cricetinae
MH  - Cricetulus
MH  - DNA/*analysis
MH  - Female
MH  - Fibroblasts/analysis/cytology
MH  - Flow Cytometry/*methods
MH  - Humans
MH  - Interphase
MH  - Leukemia L1210/analysis
MH  - Liver/cytology
MH  - Lymphocyte Activation
MH  - Lymphocytes/analysis/cytology
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Ovary/analysis/cytology
MH  - RNA/*analysis
EDAT- 1983/01/01 00:00
MHDA- 1983/01/01 00:01
CRDT- 1983/01/01 00:00
PHST- 1983/01/01 00:00 [pubmed]
PHST- 1983/01/01 00:01 [medline]
PHST- 1983/01/01 00:00 [entrez]
AID - 10.1002/cyto.990030407 [doi]
PST - ppublish
SO  - Cytometry. 1983 Jan;3(4):269-75. doi: 10.1002/cyto.990030407.