An ultra-pure in vitro phase synchrony method employing centrifugal elutriation and viable flow cytometric cell sorting.

G. C. Rice, P. N. Dean, Joe Gray, W. C. Dewey

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

We present a method of synchronizing cells in G1-, S-, and G2M-phases employing sequential centrifugal elutriation and viable flow cytometric cell sorting of Hoechst-33342 stained Chinese hamster ovary cells. G1- and S-phase cells can be separated to greater than 99% homogeneity and G2-M to 70% purity. Most of the 30% contamination in the G2-M fraction was due to S-phase cells, whose reproductive integrity could be eliminated through the use of high specific activity 3H-TdR. There were minimal toxic effects or perturbations to growth following the selection procedures. The most significant limitation of this technique appears to be the rate of cell sorting, which, with current equipment, is approximately 3,000 cells per second.

Original languageEnglish (US)
Pages (from-to)289-298
Number of pages10
JournalCytometry
Volume5
Issue number3
StatePublished - May 1984
Externally publishedYes

Fingerprint

S Phase
Poisons
G1 Phase
Cricetulus
In Vitro Techniques
Ovary
Equipment and Supplies
Growth

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Endocrinology
  • Hematology
  • Pathology and Forensic Medicine

Cite this

An ultra-pure in vitro phase synchrony method employing centrifugal elutriation and viable flow cytometric cell sorting. / Rice, G. C.; Dean, P. N.; Gray, Joe; Dewey, W. C.

In: Cytometry, Vol. 5, No. 3, 05.1984, p. 289-298.

Research output: Contribution to journalArticle

@article{7706a1a28ebc4ecaa88acdc080354767,
title = "An ultra-pure in vitro phase synchrony method employing centrifugal elutriation and viable flow cytometric cell sorting.",
abstract = "We present a method of synchronizing cells in G1-, S-, and G2M-phases employing sequential centrifugal elutriation and viable flow cytometric cell sorting of Hoechst-33342 stained Chinese hamster ovary cells. G1- and S-phase cells can be separated to greater than 99{\%} homogeneity and G2-M to 70{\%} purity. Most of the 30{\%} contamination in the G2-M fraction was due to S-phase cells, whose reproductive integrity could be eliminated through the use of high specific activity 3H-TdR. There were minimal toxic effects or perturbations to growth following the selection procedures. The most significant limitation of this technique appears to be the rate of cell sorting, which, with current equipment, is approximately 3,000 cells per second.",
author = "Rice, {G. C.} and Dean, {P. N.} and Joe Gray and Dewey, {W. C.}",
year = "1984",
month = "5",
language = "English (US)",
volume = "5",
pages = "289--298",
journal = "Communications in Clinical Cytometry",
issn = "0196-4763",
publisher = "John Wiley and Sons Inc.",
number = "3",

}

TY - JOUR

T1 - An ultra-pure in vitro phase synchrony method employing centrifugal elutriation and viable flow cytometric cell sorting.

AU - Rice, G. C.

AU - Dean, P. N.

AU - Gray, Joe

AU - Dewey, W. C.

PY - 1984/5

Y1 - 1984/5

N2 - We present a method of synchronizing cells in G1-, S-, and G2M-phases employing sequential centrifugal elutriation and viable flow cytometric cell sorting of Hoechst-33342 stained Chinese hamster ovary cells. G1- and S-phase cells can be separated to greater than 99% homogeneity and G2-M to 70% purity. Most of the 30% contamination in the G2-M fraction was due to S-phase cells, whose reproductive integrity could be eliminated through the use of high specific activity 3H-TdR. There were minimal toxic effects or perturbations to growth following the selection procedures. The most significant limitation of this technique appears to be the rate of cell sorting, which, with current equipment, is approximately 3,000 cells per second.

AB - We present a method of synchronizing cells in G1-, S-, and G2M-phases employing sequential centrifugal elutriation and viable flow cytometric cell sorting of Hoechst-33342 stained Chinese hamster ovary cells. G1- and S-phase cells can be separated to greater than 99% homogeneity and G2-M to 70% purity. Most of the 30% contamination in the G2-M fraction was due to S-phase cells, whose reproductive integrity could be eliminated through the use of high specific activity 3H-TdR. There were minimal toxic effects or perturbations to growth following the selection procedures. The most significant limitation of this technique appears to be the rate of cell sorting, which, with current equipment, is approximately 3,000 cells per second.

UR - http://www.scopus.com/inward/record.url?scp=0021433704&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0021433704&partnerID=8YFLogxK

M3 - Article

C2 - 6539665

AN - SCOPUS:0021433704

VL - 5

SP - 289

EP - 298

JO - Communications in Clinical Cytometry

JF - Communications in Clinical Cytometry

SN - 0196-4763

IS - 3

ER -