The ICCS Quality and Standards committee is dedicated to the optimization of fundamental flow cytometric testing components. Its purpose is to identify major areas of variability, determine critical components needing standardization, develop and define acceptability standards and criteria, and provide guidance and measures for practical implementation in the laboratory. This group will work closely with the ICCS education committee and other entities as necessary.
The Q&S committee is comprised of 4 groups (instrument optimization, reagents and panels, specimen preparation and reporting) which will address the most common areas of variability in flow cytometry. The information will be presented in peer-reviewed “modules” with the goal to provide the laboratory staff with a practical reference guide in optimizing their procedures.
Lysing Methods and Reagents for Flow cytometric Immunophenotyping
By Melanie O’Donahue, MT, ASCP, CCy and Laura Johnson, MT, ASCP, SM, SH
There is no consensus in the flow cytometry industry on which method of lysing erythrocytes is optimal. Different protocols might be more appropriate in different situations and differences in specimen preparation are a potential source of variability regarding the final results. In this first ICCS Quality and Standards module two experienced flow cytometry technologists present their findings about how the most commonly used lysing reagents may impact the quality of the results. We encourage you to email us at firstname.lastname@example.org with any questions you might have regarding this module.
Instrument optimization - Adjusting PMT voltages and compensation on a Beckman Coulter System
By Andrea Illingworth
Multicolor flow cytometry has evolved over the past years and has become more complex due to the number of PMT's and the associated potential for incorrect voltage and compensation settings. Instrument optimization is a much underestimated source of variability and it is important to optimize the voltages for each PMT in order to place the antigen-negative and antigen-positive population visibly "on-scale" and to maximize the potential resolution (signal/noise ratio). This is important to produce good resolution for dimly expressed antigens as well as visualization of antigen negative populations (e.g. PNH). This process should be followed at initial assay setup and this protocol can be used for QC purposes to document and track MFI and signal/noise ratio.