Indian Journal of Clinical Biochemistry

, Volume 19, Issue 2, pp 95–99 | Cite as

Enumeration of lymphocyte subsets using flow cytometry: Effect of storage before and after staining in a developing country setting

  • Sanju Jalla
  • Sunil Sazawal
  • Salkat Deb
  • robert E. Black
  • Satya Narayan Das
  • Archana Sarkar
  • Maharaj K. Bhan


Lymphocyte subset estimations by flow cytometry in population-based studies require transportation of samples from the field site to the laboratory. As samples arrive late in the day they have to wait overnight before being processed. The effect of two possible approaches, sample storage for 24 h before staining and immediate staining with analysis after 24 h and 48 h were evaluated. Two sets of experiments were performed with EDTA (ethylenediamine tetra-acetate) anticoagulated peripheral blood. In the first experiment, after collection, each sample was divided into two portions. One portion was stained at the time of blood collection and the other 24 h later after keeping it at room temperature (38–45°C). In the second experiment, blood samples were stained within 1–2 h. Each sample was analyzed immediately upon completion of staining process and subsequently after 24 h and 48 h of storage at 4°C. Results suggest that blood collected in EDTA can be processed using whole blood lysis method, after storage at room temperature (38–45°C) for 24 h with some but not significant alteration in T-cell subsets. Storage at 4°C after staining for 24 h results in a lesser and insignificant loss of cells or alteration of T-cell subsets and may be the method of choice.

Key Words

Lymphocyte storage flow cytometry leukocyte phenotypes 


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Copyright information

© Association of Clinical Biochemists of India 2004

Authors and Affiliations

  • Sanju Jalla
    • 1
  • Sunil Sazawal
    • 3
  • Salkat Deb
    • 3
  • robert E. Black
    • 2
  • Satya Narayan Das
    • 4
  • Archana Sarkar
    • 3
  • Maharaj K. Bhan
    • 5
  1. 1.Center for Cancer ResearchJohn Hopkins University School of MedicineBaltimoreUSA
  2. 2.Blumenberg School of Public HealthJohns Hopkins UniversityBaltimoreUSA
  3. 3.Center for Micronutrient ResearchAnnamalai UniversityChidambaram
  4. 4.Department of BiotechnologyAll India Institute of Medical SciencesNew Delhi
  5. 5.Department of PediatricsAll India Institute of Medical SciencesNew Delhi

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