Measurement of the cytosolic free calcium ion concentration of individual lymphocytes by microfluorometry using quin 2 or fura-2.
-
- Komada Hiroshi
- Microbiology Mie University School of Medicine
-
- Nakabayashi Hiroshi
- Departments of Patholoky Mie University School of Medicine
-
- Yoshida Toshimichi
- Departments of Patholoky Mie University School of Medicine
-
- Takanari Hideki
- Departments of Patholoky Mie University School of Medicine
説明
The cytosolic free calcium ion concentration ([Ca2+]i) of individual lymphocytes was measured by microfluorometry with dual excitation wavelengths using quin 2 or fura-2. Fura-2 was a more suitable fluorescent Ca2+ indicator than quin 2 for measurements of single cells because the standard curve calibrated for fura-2 had a good linearity, and the standard deviation (SD) of the value of the intensity ratio of fura-2-loaded cells was much smaller than that of quin 2-loaded cells. The [Ca2+]i in quiescent lymphocytes was about 1 x 10-7 M, and an increase in the [Ca2+]i was observed within a few minutes of ionomycin, protein A, phorbol myristate acetate (PMA) or concanavalin A (Con A) stimulation. Ionomycin-induced proliferation occurred when the initial [Ca2+]i was approximately 3 x 10-7 M or greater. The increase in the [Ca2+]i induced by Con A occurred transiently, and another rise in the [Ca2+]i was observed in the stage prior to the S-phase. These results indicate that Ca2+ is necessary for stimulated lymphocytes to enter the cell cycle and S-phase.
収録刊行物
-
- Cell Structure and Function
-
Cell Structure and Function 14 (2), 141-150, 1989
日本細胞生物学会
