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Evaluation of a method for counting absolute numbers of cells with a flow cytometer.

Publication ,  Journal Article
Nicholson, JK; Stein, D; Mui, T; Mack, R; Hubbard, M; Denny, T
Published in: Clin Diagn Lab Immunol
May 1997

We evaluated a method for performing absolute cell counts of lymphocyte populations with a flow cytometer. In this method, TruCount, test tubes that contain a known number of brightly fluorescent polystyrene beads are provided by the manufacturer. Whole anticoagulated blood is accurately pipetted into the tubes and mixed with fluorochrome-labeled monoclonal antibodies, the erythrocytes are lysed, and this mixture is analyzed on the flow cytometer. Absolute counts of lymphocyte subsets are calculated by determining the ratio of beads to the cell population of interest and then multiplying this ratio by the number of beads in the tube. We found this method to be reproducible. The values we obtained by the TruCount method were 5 to 10% higher than those obtained by conventional methods (flow cytometry and automated hematology) used to determine absolute numbers of cells. We believe that these differences are due to the methods of determining absolute cell counts and not to faulty identification of lymphocyte subsets.

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Published In

Clin Diagn Lab Immunol

DOI

ISSN

1071-412X

Publication Date

May 1997

Volume

4

Issue

3

Start / End Page

309 / 313

Location

United States

Related Subject Headings

  • T-Lymphocyte Subsets
  • Middle Aged
  • Microbiology
  • Infant
  • Immunology
  • Humans
  • HIV Infections
  • Fluorescent Dyes
  • Flow Cytometry
  • Evaluation Studies as Topic
 

Citation

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Nicholson, J. K., Stein, D., Mui, T., Mack, R., Hubbard, M., & Denny, T. (1997). Evaluation of a method for counting absolute numbers of cells with a flow cytometer. Clin Diagn Lab Immunol, 4(3), 309–313. https://doi.org/10.1128/cdli.4.3.309-313.1997
Nicholson, J. K., D. Stein, T. Mui, R. Mack, M. Hubbard, and T. Denny. “Evaluation of a method for counting absolute numbers of cells with a flow cytometer.Clin Diagn Lab Immunol 4, no. 3 (May 1997): 309–13. https://doi.org/10.1128/cdli.4.3.309-313.1997.
Nicholson JK, Stein D, Mui T, Mack R, Hubbard M, Denny T. Evaluation of a method for counting absolute numbers of cells with a flow cytometer. Clin Diagn Lab Immunol. 1997 May;4(3):309–13.
Nicholson, J. K., et al. “Evaluation of a method for counting absolute numbers of cells with a flow cytometer.Clin Diagn Lab Immunol, vol. 4, no. 3, May 1997, pp. 309–13. Pubmed, doi:10.1128/cdli.4.3.309-313.1997.
Nicholson JK, Stein D, Mui T, Mack R, Hubbard M, Denny T. Evaluation of a method for counting absolute numbers of cells with a flow cytometer. Clin Diagn Lab Immunol. 1997 May;4(3):309–313.

Published In

Clin Diagn Lab Immunol

DOI

ISSN

1071-412X

Publication Date

May 1997

Volume

4

Issue

3

Start / End Page

309 / 313

Location

United States

Related Subject Headings

  • T-Lymphocyte Subsets
  • Middle Aged
  • Microbiology
  • Infant
  • Immunology
  • Humans
  • HIV Infections
  • Fluorescent Dyes
  • Flow Cytometry
  • Evaluation Studies as Topic