Factors affecting cell loss in flow cytometry of mouse immune cells / 西安交通大学学报(医学版)

ABSTRACT

Objective: To explore the factors affecting cell loss in multicolor flow cytometry of mouse immune cells. Methods: C57BL/6J mice were used in the experiment. Splenocytes were isolated and resuspended twice with buffers of different types, compositions and pH; then the number of viable cells was counted to observe which factors affected the amount of cell loss during staining. Results: During flow cytometric labeling of mouse immune cells, buffer types significantly affected the amount of cell loss. Washing with PBS buffer containing potassium ions caused obvious cell loss, while slight shift in buffer pH and whether or not FBS/BSA was added had no significant effect on cell loss. Moreover, centrifugation speed also significantly affected cell loss. Conclusion: The use of potassium-containing PBS buffer during flow cytometric staining of mouse immune cells resulted in significant cell loss. The potassium-free PBS buffer and moderate centrifugation speed should be recommended.