Improved kinetic analysis of cytosolic free calcium in pressure-sensitive neuronal cells by fixed-time flow cytometry.
Two flow cytometric techniques were used to measure rapid transient changes in [Ca2+] in the neuronal cell line NH15-CA2. Using on-line injection, the cell suspension and stimulating solution are mixed and delivered to the detection point by a rapid increase in sample pressure. In NH15-CA2, injection of medium alone resulted in [Ca2+]i increase. Using the fixed-time method, where cells are maintained at constant pressure, no [Ca2+ ]i, increase was observed with medium alone. These results show that a rapid pressure increase alone alters the [Ca2+]i in NH15-CA2 cells. Both methods showed similar kinetics of [Ca2+], in response to bradykinin but the fixed-time method was found to be better for determination of the percentage of responsive cells.
Tárnok
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Center for Molecular Neurobiology, University of Hamburg, Germany.
D016475:3T3 Cells; D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D002118:Calcium / Q000032:analysis* / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology*; D045744:Cell Line, Tumor; D003470:Culture Media / Q000494:pharmacology; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism*; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D007700:Kinetics; D051379:Mice; D018613:Microscopy, Confocal; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D011312:Pressure; D011930:Reaction Time / Q000187:drug effects / Q000502:physiology; D013997:Time Factors
D003470:Culture Media; D001920:Bradykinin; D002118:Calcium