Cryo-fluorescence microscopy facilitates correlations between light and cryo-electron microscopy and reduces the rate of photobleaching.

摘要: Summary Fluorescence light microscopy (LM) has many advantages for the study of cell organization. Specimen preparation is easy and relatively inexpensive, use appropriate tags gives scientists ability to visualize specific proteins interest. LM is, however, limited in resolution, so when one interested ultrastructure, must turn electron (EM), even though this method presents problems its own. The biggest difficulty with cellular EM utility localizing macromolecules interest while retaining good structural preservation. We have built a cryo-light microscope stage that allows us generate images vitreous samples prepared cryo-EM. Correlative find areas particular by using fluorescent or vital dyes as markers within vitrified samples. Once located, sample can be placed further at higher resolution. An additional benefit cryo-LM photobleaching slower cryogenic temperatures (−140°C) than room temperature.