Protocol detail

Quantifying Intracellular GCaMP3 in Neurons

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We measured intracellular GCaMP3 concentrations in acute coronal brain slice of Emx1-Cre:Ai38 mice. The cytosolic GCaMP3 fluorescence in multiple individual neurons was measured using a 2 photon microcope. The slice was incubated in artificial cerebrospinal fluid with 20 mM extracelluar [Ca²⁺] and 1 mM ionomycin to produce saturating intracellular calcium levels. Neuronal brightness was compared to calibrated solutions of GCaMP3 in a thin cuvette.

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Zariwala H.A., Borghuis B.G., Hoogland T.M., Madisen L., Tian L., De Zeeuw C.I., Zeng H., Looger L.L., Svoboda K, & Chen T.W. (2012). A Cre-Dependent GCaMP3 Reporter Mouse for Neuronal Imaging In Vivo. The Journal of Neuroscience, 32(9), 3131-3141.

Publication 2012

Brain Calcium Cerebrospinal fluid Cytosolic Fluorescence Intracellular Ionomycin Mice Neuronal

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Protocol cited in 21 other protocols

Variable analysis

independent variables

Extracellular [Ca2+] (20 mM)
Ionomycin (1 mM)

dependent variables

Intracellular GCaMP3 concentration
Neuronal brightness

control variables

Brain slice from Emx1-Cre:Ai38 mice
Acute coronal brain slice
Artificial cerebrospinal fluid

controls

Calibrated solutions of GCaMP3 in a thin cuvette (positive control)

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