Flow cytometric measurement of calcium influx in murine T cell hybrids using Fluo-3 and an organic-anion transport inhibitor.
A method is described to facilitate flow cytometric analysis of calcium mobilization upon stimulation of murine T cell hybrids. In these transformed cell lines, the accuracy of cytometric measurement of free cytoplasmic calcium with Fluo-3 is compromised by the rapid loss of the intracellular dye. We have found that the addition of sulfinpyrazone, a known organic-anion transporter inhibitor in epithelial cells and in macrophages, severely impairs the leakage of the Fluo-3 probe from the cytoplasmic matrix. Under appropriate conditions, sulfinpyrazone has little effect on the cell physiology and permits the detection of calcium influx in a variety of murine T cell hybrids.
Baus
E
E
Laboratoire de Physiologie Animale, Université Libre de Bruxelles, Rhode Saint-Genèse, Belgium.
Urbain
J
J
Leo
O
O
Andris
F
F
D000814:Aniline Compounds; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes; D006825:Hybridomas / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D017136:Ion Transport / Q000187:drug effects; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D015398:Signal Transduction; D013442:Sulfinpyrazone / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D014966:Xanthenes
D000814:Aniline Compounds; D005456:Fluorescent Dyes; D011948:Receptors, Antigen, T-Cell; D014966:Xanthenes; C059715:Fluo-3; D002118:Calcium; D013442:Sulfinpyrazone