Comparison of flow cytometers using the side population technique to identify stem cells (TECH2P.900)

Abstract

Abstract Traditional methods using the side population analysis to identify stem cells involved the use of a flow cytometer with an ultra violet laser excited dye. This presents some challenges for researchers in relation to costs, and even though other dyes such as DyeCycle Violet (DCV) are now available that use the more economical violet laser, the cost of a flow cytometer to provide this type of analysis is still an obstacle. More recently, new benchtop flow cytometers have become available that can provide the quality of a high-end flow cytometer at a much more affordable cost. In this analysis, we sought out to identify stem cells and progenitors using the side population technique in murine bone marrow using the ACEA NovoCyte flow cytometer which maximizes optical design through its use of shared PMTs. Samples were stained and evaluated with (DCV), Lineage markers, Sca, and c-kit. To identify stem cells using the side population technique, we first gated on Lineage- cells and subsequently on Sca+, c-kit+. To confirm the correct identification of these cells, this population was back gated comparing DCV red fluorescence vs DCV blue fluorescence and verified the cells are localized to the expected tail-region. This study is evidence that quality data can be generated on a budget-friendly flow cytometer and allows researchers with limited resources to use flow cytometry.