Analysis of free intracellular calcium by flow cytometry: multiparameter and pharmacologic applications.
Flow cytometry offers numerous advantages over traditional techniques for measuring intracellular Ca(2+) in lymphoid and nonlymphoid cells. In particular, the heterogeneity of cell responses can be defined by flow cytometry, and multiparameter analyses permit the determination of intracellular Ca(2+) in surface-marker-defined target cells as well as correlation of changes in Ca(2+) with other biochemical markers, including ligand binding. This article presents several established methods for measuring intracellular Ca(2+) by flow cytometry in lymphoid and nonlymphoid cells. Examples are provided for determination of Ca(2+) in human peripheral blood leukocytes and two human epithelial cell lines grown in monolayer. In addition, applications are reviewed or presented for correlating changes in intracellular Ca(2+) with other cell parameters, including cell cycle analysis, changes in cell membrane integrity, and the induction of apoptosis markers. Finally, a number of novel sample handling capabilities useful for performing kinetic analyses of Ca(2+) changes by flow cytometry are now available and one application is presented which is finding utility in pharmacologic studies.
Burchiel
S W
SW
College of Pharmacy, University of New Mexico, Albuquerque, New Mexico, 87131, USA.
Edwards
B S
BS
Kuckuck
F W
FW
Lauer
F T
FT
Prossnitz
E R
ER
Ransom
J T
JT
Sklar
L A
LA
D000814:Aniline Compounds / Q000378:metabolism; D001572:Benzofurans / Q000378:metabolism; D002118:Calcium / Q000032:analysis* / Q000378:metabolism*; D002138:Calibration; D002448:Cell Adhesion; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002614:Chelating Agents / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007093:Imidazoles / Q000378:metabolism; D007424:Intracellular Fluid / Q000737:chemistry* / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism; D011419:Propidium / Q000378:metabolism; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000235:genetics / Q000378:metabolism; D018000:Receptors, Peptide / Q000235:genetics / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014162:Transfection; D014966:Xanthenes / Q000378:metabolism
D000814:Aniline Compounds; D001572:Benzofurans; D002614:Chelating Agents; D005456:Fluorescent Dyes; D007093:Imidazoles; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; D018000:Receptors, Peptide; D014966:Xanthenes; C082877:fura red; C059715:Fluo-3; D011419:Propidium; D002118:Calcium