A novel method for kinetic measurements of rare immune cell proliferation using Cellometer image-based cytometry (124.1)

Abstract

Abstract Measuring cell proliferation is important for studying the effects of various treatments on cultured primary cells or cell lines. Current proliferation analysis methods employ flow cytometry for fluorescence detection of CFSE-labeled cells. However, conventional flow cytometers require a considerable amount of cells per reading, which becomes an issue for kinetic measurements with rare cell population due to the lack of samples for flow cytometric analyses at multiple time points during proliferation period. Here we report the development of a novel cell proliferation kinetic detection method for low cell concentration samples using the new Cellometer Vision image-based cytometry system. Since the Cellometer system requires only 20 µl of sample, cell proliferation can be measured at multiple time points over the proliferation period, whereas typically, flow cytometry is only performed at the end of the proliferation period. To validate the detection method, B1 and B2 B cells were treated with a B cell mitogen and proliferation was measured on day 1, 3, 5, and 6. To demonstrate the capability, B1 B cells were treated with a panel of TLR agonists and proliferation was measured on day 2, 4, 6, and 7. Cellometer was able to obtain proliferation results on each day, which were comparable to flow cytometry. This novel method allows for kinetic measurements of the rare cell samples such as B1 B cell, which has the potential to revolutionize kinetic analysis of cell proliferation.