Open AccessImproved kinetic analysis of cytosolic free calcium in pressure‐sensitive neuronal cells by fixed‐time flow cytometry
Author(s) -
Tárnok Attila
Publication year - 1996
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(19960101)23:1<82::aid-cyto13>3.0.co;2-s
Subject(s) - flow cytometry , calcium , cytometry , biophysics , cytosol , chemistry , flow (mathematics) , stopped flow , microbiology and biotechnology , kinetics , materials science , biology , biochemistry , physics , enzyme , mechanics , organic chemistry , quantum mechanics , reaction rate constant
Two flow cytometric techniques were used to measure rapid transient changes in [Ca 2+ ] i 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 [Ca 2+ ] i increase. Using the fixed‐time method, where cells are maintained at constant pressure, no [Ca 2+ ] i increase was observed with medium alone. These results show that a rapid pressure increase alone alters the [Ca 2+ ] i in NH15‐CA2 cells. Both methods showed similar kinetics of [Ca 2+ ] i in response to bradykinin but the fixed‐time method was found to be better for determination of the percentage of responsive cells. © 1996 Wiley‐Liss, Inc.