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Flow Cytometry01:23

Flow Cytometry

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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Analyzing Platelet Subpopulations by Multi-color Flow Cytometry
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Platelet function investigation by flow cytometry: Sample volume, needle size, and reference intervals.

Oliver Heidmann Pedersen1, Peter H Nissen1, Anne-Mette Hvas1

  • 1a Centre for Haemophilia and Thrombosis, Department of Clinical Biochemistry , Aarhus University Hospital , Aarhus , Denmark.

Platelets
|September 30, 2017
PubMed
Summary
This summary is machine-generated.

Flow cytometry for platelet function analysis can now use small blood volumes (1.0 mL) and standard needles without affecting results. This study establishes reference intervals for this vital diagnostic tool.

Keywords:
Blood samplingflow cytometryplatelet function analysisplateletsreference interval

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Area of Science:

  • Hematology
  • Immunology
  • Analytical Chemistry

Background:

  • Flow cytometry offers advantages for platelet function analysis, including small sample volume requirements.
  • Expanding applications, such as pediatric testing, necessitates even smaller blood volumes.
  • Standardized reference intervals are crucial for clinical adoption of flow cytometric platelet function assays.

Purpose of the Study:

  • To assess the feasibility of using reduced whole blood volumes (1.0 mL) for flow cytometric platelet function analysis.
  • To investigate the impact of needle gauge (19- vs. 21-gauge) on platelet pre-activation during blood sampling.
  • To establish reference intervals for key platelet activation markers using flow cytometry.

Main Methods:

  • Blood samples from healthy individuals were analyzed using varying volumes (1.0, 1.8, 3.5 mL) and needle sizes (19, 21 gauge).
  • Platelet activation markers (bound-fibrinogen, CD63, P-selectin/CD62p) were measured via flow cytometry after stimulation with agonists.
  • Reference intervals were determined from 78 healthy adults using a NAVIOS flow cytometer.

Main Results:

  • Flow cytometric platelet function analysis is reliable with blood volumes as low as 1.0 mL.
  • Both 19-gauge and 21-gauge needles are suitable for blood collection without causing significant platelet pre-activation.
  • Reference intervals for flow cytometric platelet function assays were successfully established.

Conclusions:

  • Flow cytometry is a robust method for platelet function testing, adaptable to smaller sample volumes.
  • Standard blood collection techniques are compatible with flow cytometric analysis, ensuring reliable results.
  • Established reference intervals will facilitate the clinical implementation of flow cytometry for platelet function assessment.