Abstract:
:Glial cells have been identified as key signaling components in the brain; however, methods to investigate their structure and function in vivo have been lacking. Here, we describe a new, highly selective approach for labeling astrocytes in intact rodent neocortex that allows in vivo imaging using two-photon microscopy. The red fluorescent dye sulforhodamine 101 (SR101) was specifically taken up by protoplasmic astrocytes after brief exposure to the brain surface. Specificity was confirmed by immunohistochemistry. In addition, SR101 labeled enhanced green fluorescent protein (EGFP)-expressing astrocytes but not microglial cells in transgenic mice. We used SR101 labeling to quantify morphological characteristics of astrocytes and to visualize their close association with the cortical microvasculature. Furthermore, by combining this method with calcium indicator loading of cell populations, we demonstrated distinct calcium dynamics in astroglial and neuronal networks. We expect SR101 staining to become a principal tool for investigating astroglia in vivo.
journal_name
Nat Methodsjournal_title
Nature methodsauthors
Nimmerjahn A,Kirchhoff F,Kerr JN,Helmchen Fdoi
10.1038/nmeth706keywords:
subject
Has Abstractpub_date
2004-10-01 00:00:00pages
31-7issue
1eissn
1548-7091issn
1548-7105pii
nmeth706journal_volume
1pub_type
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