Preferential Loading of Bergmann Glia with Synthetic Acetoxymethyl Calcium Dyes
Adapted from Imaging in Neuroscience (ed. Helmchen and Konnerth). CSHL Press, Cold Spring Harbor, NY, USA, 2011.Abstract
The cerebellar cortex contains two astrocyte types: the Bergmann glia of the molecular layer and the velate protoplasmic astrocytes of the granule cell layer. In vivo, these cell types generate both subcellular calcium transients and trans-glial calcium waves. This protocol outlines a method for in vivo calcium imaging in cerebellar astrocytes of mice which have undergone a cerebellar craniotomy. Multicell bolus loading (MCBL) is performed using the synthetic calcium indicators Fluo-5F AM and Fluo-4 AM. In the cerebellum, a degree of cell-type specificity can be achieved by varying the depth of injection. This protocol describes a loading procedure following craniotomy which allows preferential labeling of Bergmann glia.
- © 2011 Cold Spring Harbor Laboratory Press
