Abstract
Calcium imaging is a technique in which Ca2+-binding molecules are loaded into live cells and as they bind Ca2+ they “indicate” the concentration of free calcium through a change in either the intensity or the wavelength of light emitted (fluorescence or bioluminescence). There are several possible methods for loading synthetic Ca2+ indicators into subcellular compartments, including topical application of membrane-permeant Ca2+ indicators, forward-filling of dextran conjugates, and direct injection. Calcium imaging is a highly informative technique in neurobiology because Ca2+ is involved in many neuronal signaling pathways and serves as the trigger for neurotransmitter release. This article describes the forward-filling of dextran-conjugated indicators at the Drosophila larval neuromuscular junction (NMJ). This technique is particularly well suited for imaging changes in cytosolic Ca2+ as dextran conjugation prevents compartmentalization of the Ca2+ indicator. The major drawback is that the nerves must be severed at the start of the loading process, several hours before nerve terminals are ready to examine.
Publisher
Cold Spring Harbor Laboratory
Subject
General Biochemistry, Genetics and Molecular Biology
Cited by
15 articles.
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