Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (24): 4481-4483.doi: 10.3969/j.issn.1673-8225.2010.24.025

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Real-time monitoring of intracellular Ca 2+ concentration by flow cytometry

Cao Yun-xin1, Li Yu-rong2, Yang An-gang1   

  1. 1 Department of Immunology, 2 Department of Preventive Medicine, the Fourth Military Medical University, Xi’an  710032, Shaanxi Province, China
  • Online:2010-06-11 Published:2010-06-11
  • About author:Cao Yun-xin, Senior Experimentalist, Department of Immunology, the Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China Caoxin87@fmmu.edu.cn
  • Supported by:

    the National Natural Science Foundation of China, No. 30700730*

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Abstract:

BACKGROUND: The concentration changes of Ca 2+ in cells is susceptible to various signal transduction pathways, thus, it is helpful to monitor changes in Ca 2+ for a better understanding of the priming, augmentation or inhibition of the functions of T-cells.
OBJECTIVE: To detect dynamic changes in Ca 2+ using flow cytometry.
METHODS: The fresh whole blood was obtained from human, and the lymphocytes were separated. CD3mAb and CD28mAb were added into the Fluo-3/AM-loaded cells, and the changes of Ca 2+ concentration were real-time monitored by flow cytometry.
RESULTS AND CONCLUSION: The fluorescence value of Ca 2+ was stable and showed a baseline when the T lymphocytes at static status; after adding anti-CD3mAb and anti-CD28mAb, the T lymphocytes were activated, and the Ca 2+ concentration was quickly increased, lasted for 2-3 minutes, and gradually trended to be stale. The results revealed that flow cytometric analysis enabled us to trace dynamic movement of Ca 2+ using Fluo-3, which can be used as an effective method for monitoring Ca 2+ changes. 

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