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Sample Preparation for Mass Cytometry Analysis
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Relevance of sample preparation for flow cytometry.

V E Muccio1, E Saraci1, M Gilestro1

  • 1Divisione Universitaria di Ematologia, A.O.U. Città della Salute e della Scienza di Torino, Torino, Italy.

International Journal of Laboratory Hematology
|October 7, 2017
PubMed
Summary

The lyse-wash-stain-wash (LWSW) method for flow cytometry sample preparation is comparable to the standard stain-lyse-wash (SLW) method for hematological disease diagnosis and minimal residual disease (MRD) studies.

Keywords:
antibody bindingflow cytometrylyse-wash-stainsample preparationstain-lyse-wash

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Area of Science:

  • Hematology
  • Immunology
  • Cell Biology

Background:

  • Flow cytometry is crucial for diagnosing hematological diseases and assessing minimal residual disease (MRD).
  • Standard sample preparation involves stain-lyse-wash (SLW).
  • Lyse-wash-stain-wash (LWSW) is proposed for enhanced sensitivity and reduced nonspecific binding in MRD studies.

Purpose of the Study:

  • To compare the efficacy of LWSW and SLW sample preparation methods.
  • To evaluate differences in mean fluorescence intensity (MFI), stain index, signal-to-noise ratio, and positive cell percentages.

Main Methods:

  • Simultaneous preparation of 10 samples using both LWSW and SLW protocols.
  • Analysis of 104 antibodies and 13 selected antibodies for key flow cytometry metrics.

Main Results:

  • MFI and positive cell percentages showed high correlation and no significant differences between LWSW and SLW.
  • LWSW demonstrated higher stain index and signal-to-noise ratios for kappa and lambda surface chains.
  • SLW showed a trend towards higher values for other antibodies.

Conclusions:

  • LWSW is a viable alternative to SLW for diagnosis and MRD evaluation, offering comparable results.
  • LWSW can prevent nonspecific binding and reduce cell debris acquisition.
  • LWSW allows for more consistent antibody intensity expression when comparing samples processed with bulk lysis.