A Novel Method for Kinetic Measurements of Rare Immune Cell Proliferation using Cellometer Image-Based Cytometry

Leo L. Chan, Xuemei Zhong, Alnoor Pirani, Benjamin Paradis, and Bo Lin

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 CFSElabeled 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 sufficient samples
to test at multiple time points during the 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 imagebased 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 also 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 that 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.