학술논문
Cellular, subcellular and functional in vivo labeling of the spinal cord using vital dyes
Document Type
Report
Author
Source
Nature Protocols. March 1, 2013, Vol. 8 Issue 3, p481, 10 p.
Subject
Language
English
ISSN
1754-2189
Abstract
INTRODUCTION In vivo microscopy is a powerful technique for visualizing cellular behavior in the healthy and diseased nervous system (1,2). In the spinal cord, for example, in vivo microscopy has [...]
Here we provide a protocol for rapidly labeling different cell types, distinct subcellular compartments and key injury mediators in the spinal cord of living mice. This method is based on the application of synthetic vital dyes to the surgically exposed spinal cord. Suitable vital dyes applied in appropriate concentrations lead to reliable in vivo labeling, which can be combined with genetic tags and in many cases preserved for postfixation analysis. In combination with in vivo imaging, this approach allows the direct observation of central nervous system physiology and pathophysiology at the cellular, subcellular and functional level. Surgical exposure and preparation of the spinal cord can be achieved in less than 1 h, and then dyes need to be applied for 30- 60 min before the labeled spinal cord can be imaged for several hours.
Here we provide a protocol for rapidly labeling different cell types, distinct subcellular compartments and key injury mediators in the spinal cord of living mice. This method is based on the application of synthetic vital dyes to the surgically exposed spinal cord. Suitable vital dyes applied in appropriate concentrations lead to reliable in vivo labeling, which can be combined with genetic tags and in many cases preserved for postfixation analysis. In combination with in vivo imaging, this approach allows the direct observation of central nervous system physiology and pathophysiology at the cellular, subcellular and functional level. Surgical exposure and preparation of the spinal cord can be achieved in less than 1 h, and then dyes need to be applied for 30- 60 min before the labeled spinal cord can be imaged for several hours.