Visualizing calcium signaling in cells by digitized wide-field and confocal fluorescent microscopy.
Calcium (Ca2+) is a fundamentally important component of cellular signal transduction. Dynamic changes in the concentration of Ca2+ ([Ca2+]) in the cytoplasm and within organelles are tightly controlled and regulate a diverse array of biological activities, including fertilization, cell division, gene expression, cellular metabolism, protein biosynthesis, secretion, muscle contraction, intercellular communication, and cell death. Measurement of intracellular [Ca2+] is essential to understanding the role of Ca2+ and for defining the underlying regulatory mechanisms in any cellular process. A broad range of synthetic and biosynthetic fluorescent Ca2+ sensors are available that enable the visualization and quantification of subcellular spatio-temporal [Ca2+] gradients. This chapter describes the application of wide-field digitized video fluorescence microfluorometry and confocal microscopy to quantitatively image Ca2+ in cells with high temporal and spatial resolution.
Roe
Michael W
MW
Department of Medicine, University of Chicago, IL, USA.
Fiekers
Jerome F
JF
Philipson
Louis H
LH
Bindokas
Vytautas P
VP
D000818:Animals; D015374:Biosensing Techniques; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling / Q000502:physiology*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D005456:Fluorescent Dyes / Q000378:metabolism*; D051379:Mice; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure
D005456:Fluorescent Dyes; D002118:Calcium