Document Detail

Cell flow cytometry of fixed and unfixed bursal and thymic cells.
MedLine Citation:
PMID:  4001057     Owner:  NLM     Status:  MEDLINE    
Analysis in a cell flow cytometer (Ortho Spectrum III, Ortho Inst.) of single cell suspensions from the bursa and thymus of 20-day embryos revealed two distinct cell clusters. The two clusters were less apparent after fixation of the cells in paraformaldehyde and assumed a comet-like appearance at 30 min fixation in ethyl alcohol (EA). The G (postmitotic), S [deoxyribonucleic acid (DNA) synthesis], and G2/M (premitotic and mitotic) phases of the life cycle were visualized in two cell flow cytometers (Ortho Spectrum III and FACS IV, Bectin Dickinson) after treating the cells with EA, ribonuclease (RNase), and propidium iodide (PI, a fluorescent dye). Bursal cell suspensions exposed to the EA-RNase-PI protocol and stored for 3 weeks in phosphate-buffered saline showed minor changes in the G1 coefficient of variation, G1, and S percentages, but marked changes in these parameters occurred after 6 weeks of storage. Thymic cells treated in a similar fashion could not be maintained for 3 weeks. Bursal and thymic cells may remain in the EA for one day and possibly as long as 7 days prior to preparing them for DNA life cycle analysis. Paraformaldehyde was not a satisfactory cell fixative for assessing a cell's DNA life cycle.
B Glick; M F LaVia; B Koger
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Poultry science     Volume:  64     ISSN:  0032-5791     ISO Abbreviation:  Poult. Sci.     Publication Date:  1985 Apr 
Date Detail:
Created Date:  1985-07-25     Completed Date:  1985-07-25     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  0401150     Medline TA:  Poult Sci     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  737-43     Citation Subset:  IM    
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MeSH Terms
Bursa of Fabricius / cytology*,  embryology
Cell Cycle
Chick Embryo / cytology*
DNA / analysis
Ethanol / pharmacology
Flow Cytometry
Formaldehyde / pharmacology
Lymphocytes / analysis,  cytology*,  drug effects
Polymers / pharmacology
Thymus Gland / cytology*,  embryology
Grant Support
1F33A106703//PHS HHS
Reg. No./Substance:
0/Fixatives; 0/Polymers; 30525-89-4/paraform; 50-00-0/Formaldehyde; 64-17-5/Ethanol; 9007-49-2/DNA

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