| First Author | Verma M | Year | 2011 |
| Journal | J Histochem Cytochem | Volume | 59 |
| Issue | 1 | Pages | 60-7 |
| PubMed ID | 20876523 | Mgi Jnum | J:170193 |
| Mgi Id | MGI:4944128 | Doi | 10.1369/jhc.2010.956730 |
| Citation | Verma M, et al. (2011) Efficient single muscle fiber isolation from alcohol-fixed adult muscle following beta-galactosidase staining for satellite cell detection. J Histochem Cytochem 59(1):60-7 |
| abstractText | Staining for beta-galactosidase activity for whole tissues, sections, and cells is a common method to detect expression of beta-galactosidase reporter transgene as well as senescence-dependent beta-galactosidase activity. Choice of fixatives is a critical step for detection of beta-galactosidase activity, subsequent immunostaining, and enzymatic digestion of tissue to dissociate cells. In this report, the authors examined several aldehyde and alcohol fixatives in mouse skeletal muscle tissues for their efficiency at improving detection of beta-galactosidase activity as well as detection by immunostaining. In addition, fixatives were also analyzed for their efficiency for collagenase digestion to isolate single muscle fibers on postfixed beta-galactosidase-stained whole skeletal muscle tissues. The results show that fixing cells with isopropanol yields the greatest reliability and intensity in both beta-galactosidase staining as well as double staining for beta-galactosidase activity and antibodies. In addition, isopropanol and ethanol, but not glutaraldehyde or paraformaldehyde, allow for the isolation of single muscle fibers from the diaphragm and tibialis anterior muscles following postfixed beta-galactosidase staining. Using this method, it is possible to identify the amount of cells that occupy the satellite cell compartment in single muscle fibers prepared from any muscle tissues, including tibialis anterior muscle and diaphragm. |
