Journal of Histochemistry and Cytochemistry Priciples for Free Access to Science
  Search:   
    >> Advanced Search

Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Deitch, A. D.
Right arrow Articles by deVere White, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Deitch, A. D.
Right arrow Articles by deVere White, R.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

A stable propidium iodide staining procedure for flow cytometry

AD Deitch, H Law and R deVere White

A propidium iodide (PI) staining procedure is described in which 50 micrograms/ml PI in 10(-2) M Tris, pH 7.0, with 5 mM MgCl2 is used to stain murine erythroleukemia cells (MELC) grown in suspension culture as well as single cell suspensions derived from rat kidney adenocarcinoma and human prostatic carcinoma. Specificity of staining of nuclear DNA is achieved by enzymatic removal of RNA using RNAse in the staining solution. Virtually identical histograms, with the same G1 peak height and closely similar coefficients of variation (CVs), are obtained using a wide range of RNAse concentrations on replicate samples of MELC if the incubation times are sufficiently prolonged when employing the lower enzyme concentrations. For 1 mg/ml RNAse on logarithmically growing MELC, 30 min incubation at 37 degrees C is needed to obtain a maximum G1 peak height and optimal CV and there is no significant change in the histogram if the incubation is prolonged to 4 hr. For every 4-fold decrease in RNAse concentration, the incubation time at 37 degrees C must be doubled to obtain the same maximal G1 peak height and optimal CV. Unfixed cell preparations, whether derived from suspension or monolayer cultures or from solid tumors, are stable for 2 or more weeks if stored at 4 degrees C between flow cytometric analyses and histograms are usually only minimally altered if the stained cell samples are stored for 1-2 months at 4 degrees C. Sample decay is associated with bacterial contamination. If sterile preparative techniques are used initially, subsequent contamination of the stained preparations may be minimized by adding sodium azide to the stained samples at 0.1% without influencing fluorescence intensity. Glycerine may be added to 10% and the samples slowly frozen for storage without altering DNA histogram shapes. The simplicity of sample preparation and the stability of the resulting stained cell samples makes this procedure suitable for repetitive comparative sampling of tissue and cell populations over prolonged time spans.

Volume 30, Issue 9, pp. 967-972, 09/01/1982
Copyright © 1982 by The Histochemical Society


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
L. Amazit, Y. Alj, R. K. Tyagi, A. Chauchereau, H. Loosfelt, C. Pichon, J. Pantel, E. Foulon-Guinchard, P. Leclerc, E. Milgrom, et al.
Subcellular Localization and Mechanisms of Nucleocytoplasmic Trafficking of Steroid Receptor Coactivator-1
J. Biol. Chem., August 22, 2003; 278(34): 32195 - 32203.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Sci.Home page
M Karbowski, J. Spodnik, M Teranishi, M Wozniak, Y Nishizawa, J Usukura, and T Wakabayashi
Opposite effects of microtubule-stabilizing and microtubule-destabilizing drugs on biogenesis of mitochondria in mammalian cells
J. Cell Sci., January 1, 2001; 114(2): 281 - 291.
[Abstract] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
A. Quaroni, J. Q. Tian, P. Seth, and C. Ap Rhys
p27Kip1 is an inducer of intestinal epithelial cell differentiation
Am J Physiol Cell Physiol, October 1, 2000; 279(4): C1045 - C1057.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
H. C. Ha, A. Thiagalingam, B. D. Nelkin, and R. A. Casero Jr.
Reactive Oxygen Species Are Critical for the Growth and Differentiation of Medullary Thyroid Carcinoma Cells
Clin. Cancer Res., September 1, 2000; 6(9): 3783 - 3787.
[Abstract] [Full Text]

Home page
 Clin. Cancer Res.Home page
P. C. Mack, D. R. Gandara, C. Bowen, M. J. Edelman, T. Paglieroni, J. B. Schnier, E. P. Gelmann, and P. H. Gumerlock
RB Status as a Determinant of Response to UCN-01 in Non-Small Cell Lung Carcinoma
Clin. Cancer Res., September 1, 1999; 5(9): 2596 - 2604.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
H. C. Ha, P. M. Woster, J. D. Yager, and R. A. Casero Jr.
The role of polyamine catabolism in polyamine analogue-induced programmed cell death
PNAS, October 14, 1997; 94(21): 11557 - 11562.
[Abstract] [Full Text] [PDF]

Home page
 ScienceHome page
D. W. GALBRAITH, K. R. HARKINS, J. M. MADDOX, N. M. AYRES, D. P. SHARMA, and E. FIROOZABADY
Rapid Flow Cytometric Analysis of the Cell Cycle in Intact Plant Tissues
Science, June 3, 1983; 220(4601): 1049 - 1051.
[Abstract] [PDF]



Guidelines | Subscriptions | About | exPRESS - Current - Archive | Business Information | Contact
The Journal of Histochemistry & Cytochemistry is owned, published, and licensed by The Histochemical Society © 1982