title: Techniques for measuring and manipulating free Ca2+ in the cytosol and organelles of neutrophils.
abstract: Ca(2+) signalling in neutrophils is important for triggering and coordinating the behaviour of neutrophils. Fluorescent probes for cytosolic free Ca(2+) concentration, e.g., fura2 and fluo3, have been widely used in neutrophils. These probes can be used to monitor Ca(2+) in the cytosol, the nucleus, near the plasma membrane and theoretically within Ca(2+) storage organelles. The longer wavelength indicators, e.g., fluo3 and calcium green, can be used confocally to monitor subcellular Ca(2+) changes in the cytosol of neutrophils and in the nucleus. Confocal techniques also permit "impossible views" imaging of Ca(2+) and newer scanning techniques promise very fast temporal resolution. Techniques using chlortetracycline (CTC) and DiOC(6)(3) are also described for monitoring the position of Ca(2+) storage sites in neutrophils and for manipulating their activity. Thus, in this review, a spectrum of new (and older) optical techniques are presented which are useful for measuring, monitoring and manipulating cytosolic free Ca(2+) concentration and Ca(2+) storage in neutrophils. With these techniques, it is hoped that more insight will be gained into both the mechanism of and the consequences of Ca(2+) signalling in neutrophils.
authors.affiliation: Molecular Signalling Group, University Department of Surgery, University of Wales College of Medicine, Heath Park, Cardiff, UK. hallettmb@cardiff.ac.uk
authors.forename: M B
authors.initials: MB
authors.lastname: Hallett
authors.forename: R
authors.initials: R
authors.lastname: Hodges
authors.forename: M
authors.initials: M
authors.lastname: Cadman
authors.forename: H
authors.initials: H
authors.lastname: Blanchfield
authors.forename: S
authors.initials: S
authors.lastname: Dewitt
authors.forename: E J
authors.initials: EJ
authors.lastname: Pettit
authors.forename: I
authors.initials: I
authors.lastname: Laffafian
authors.forename: E V
authors.initials: EV
authors.lastname: Davies
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008845:Microinjections; D018613:Microscopy, Confocal; D015336:Molecular Probe Techniques; D009504:Neutrophils / Q000737:chemistry* / Q000378:metabolism; D015388:Organelles / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002118:Calcium