Journal
JOURNAL OF MICROSCOPY
Volume 227, Issue 2, Pages 98-109Publisher
WILEY
DOI: 10.1111/j.1365-2818.2007.01794.x
Keywords
green fluorescent protein; correlative light and electron microscopy; cryo-electron tomography; cryo-light microscopy; microtubules
Categories
Funding
- NCRR NIH HHS [RR000592] Funding Source: Medline
Ask authors/readers for more resources
Fluorescence light microscopy (LM) has many advantages for the study of cell organization. Specimen preparation is easy and relatively inexpensive, and the use of appropriate tags gives scientists the ability to visualize specific proteins of interest. LM is, however, limited in resolution, so when one is interested in ultrastructure, one must turn to electron microscopy (EM), even though this method presents problems of its own. The biggest difficulty with cellular EM is its limited utility in localizing macromolecules of interest while retaining good structural preservation. We have built a cryo-light microscope stage that allows us to generate LM images of vitreous samples prepared for cryo-EM. Correlative LM and EM allows one to find areas of particular interest by using fluorescent proteins or vital dyes as markers within vitrified samples. Once located, the sample can be placed in the EM for further study at higher resolution. An additional benefit of the cryo-LM stage is that photobleaching is slower at cryogenic temperatures (- 140 degrees C) than at room temperature.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available