title: Measurement of intracellular Ca2+ concentration in single cells using ratiometric calcium dyes.
abstract: Measurement of intracellular Ca(2+) concentration ([Ca(2+)](i)) is useful to study the upstream and downstream events of Ca(2+) signaling. Ca(2+)-binding proteins including EF-hand-containing proteins are important downstream effector molecules after an increase of [Ca(2+)](i). Conversely, these proteins can also act as key modulators for regulation of [Ca(2+)](i) by sensing the Ca(2+) levels in the intracellular organelles and cytoplasm. Here we describe a single-cell Ca(2+) imaging technique that was used to measure the intracellular Ca(2+) levels to examine the function of Ca(2+)-binding proteins, STIM1 and Calcium release-activated Calcium channel regulator 2A (CRACR2A), using ratiometric Ca(2+) dye Fura-2 in adherent and non-adherent cells.
authors.affiliation: Department of Physiology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA.
authors.forename: Sonal
authors.initials: S
authors.lastname: Srikanth
authors.forename: Yousang
authors.initials: Y
authors.lastname: Gwack
mesh_terms: D002118:Calcium / Q000032:analysis* / Q000378:metabolism; D002138:Calibration; D002448:Cell Adhesion; D002460:Cell Line; D004396:Coloring Agents / Q000032:analysis* / Q000378:metabolism; D016257:Fura-2 / Q000378:metabolism; D055785:Gene Knockdown Techniques; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism*; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*; D014162:Transfection
chemical_list: D004396:Coloring Agents; D034741:RNA, Small Interfering; D002118:Calcium; D016257:Fura-2