Abstract
Background: In an accompanying paper (Asmuth et al.) it was found necessary to include cell death explicitly to estimate parameters of cell proliferation. The use of bivariate flow cytometry to estimate the phase durations and the doubling times of cells labeled with thymidine analogues is well established. However, these methods of analysis do not consider the possibility of cell death. This report demonstrates that estimating cell death in G2/M is possible. Methods: Mathematical models for the experimental quantities, the fraction of labeled undivided cells, the fraction of labeled divided cells, and the relative movement were developed. These models include the possibility that, of the cells with G2/M DNA content, only a certain fraction will divide, with the remainder dying after some time TR. Simulation studies were conducted to test the possibility of using simple methods to estimate phase durations and cell death rates. Results: Cell death alters the estimates of phase transit times in a rather complex manner that depends on the lifetime of the doomed cells. However, it is still possible to obtain estimates of the phase durations of cells in S and G2/M and the death rates of cells in G2/M. Conclusions: The methods presented herein provide a new way to characterize cell populations that includes cell death rates and common measurements of cell proliferation.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 32-40 |
| Number of pages | 9 |
| Journal | Cytometry Part A |
| Volume | 66 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jul 2005 |
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Keywords
- Bromodeoxyuridine
- Cell cycle
- Phase durations
- Population growth
ASJC Scopus subject areas
- Hematology
- Cell Biology
- Pathology and Forensic Medicine
- Biophysics
- Endocrinology
Cite this
Estimating cell death in G2M using bivariate BrdUrd/DNA flow cytometry. / White, R. Allen; Asmuth, David; Lu, Ying; Wang, Nan; Li, Xiao Dong; Reece, Lisa; Pollard, Richard B; Nokta, Mostafa; Leary, James F.; Terry, Nicholas H A.
In: Cytometry Part A, Vol. 66, No. 1, 07.2005, p. 32-40.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Estimating cell death in G2M using bivariate BrdUrd/DNA flow cytometry
AU - White, R. Allen
AU - Asmuth, David
AU - Lu, Ying
AU - Wang, Nan
AU - Li, Xiao Dong
AU - Reece, Lisa
AU - Pollard, Richard B
AU - Nokta, Mostafa
AU - Leary, James F.
AU - Terry, Nicholas H A
PY - 2005/7
Y1 - 2005/7
N2 - Background: In an accompanying paper (Asmuth et al.) it was found necessary to include cell death explicitly to estimate parameters of cell proliferation. The use of bivariate flow cytometry to estimate the phase durations and the doubling times of cells labeled with thymidine analogues is well established. However, these methods of analysis do not consider the possibility of cell death. This report demonstrates that estimating cell death in G2/M is possible. Methods: Mathematical models for the experimental quantities, the fraction of labeled undivided cells, the fraction of labeled divided cells, and the relative movement were developed. These models include the possibility that, of the cells with G2/M DNA content, only a certain fraction will divide, with the remainder dying after some time TR. Simulation studies were conducted to test the possibility of using simple methods to estimate phase durations and cell death rates. Results: Cell death alters the estimates of phase transit times in a rather complex manner that depends on the lifetime of the doomed cells. However, it is still possible to obtain estimates of the phase durations of cells in S and G2/M and the death rates of cells in G2/M. Conclusions: The methods presented herein provide a new way to characterize cell populations that includes cell death rates and common measurements of cell proliferation.
AB - Background: In an accompanying paper (Asmuth et al.) it was found necessary to include cell death explicitly to estimate parameters of cell proliferation. The use of bivariate flow cytometry to estimate the phase durations and the doubling times of cells labeled with thymidine analogues is well established. However, these methods of analysis do not consider the possibility of cell death. This report demonstrates that estimating cell death in G2/M is possible. Methods: Mathematical models for the experimental quantities, the fraction of labeled undivided cells, the fraction of labeled divided cells, and the relative movement were developed. These models include the possibility that, of the cells with G2/M DNA content, only a certain fraction will divide, with the remainder dying after some time TR. Simulation studies were conducted to test the possibility of using simple methods to estimate phase durations and cell death rates. Results: Cell death alters the estimates of phase transit times in a rather complex manner that depends on the lifetime of the doomed cells. However, it is still possible to obtain estimates of the phase durations of cells in S and G2/M and the death rates of cells in G2/M. Conclusions: The methods presented herein provide a new way to characterize cell populations that includes cell death rates and common measurements of cell proliferation.
KW - Bromodeoxyuridine
KW - Cell cycle
KW - Phase durations
KW - Population growth
UR - http://www.scopus.com/inward/record.url?scp=20944436936&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=20944436936&partnerID=8YFLogxK
U2 - 10.1002/cyto.a.20147
DO - 10.1002/cyto.a.20147
M3 - Article
C2 - 15915505
AN - SCOPUS:20944436936
VL - 66
SP - 32
EP - 40
JO - Cytometry. Part A : the journal of the International Society for Analytical Cytology
JF - Cytometry. Part A : the journal of the International Society for Analytical Cytology
SN - 1552-4922
IS - 1
ER -