A correlative light and electron microscopy method based on laser micropatterning and etching.
Correlative microscopy is a hybrid method that allows the localization of events observed under visible, ultraviolet, or infrared light, at molecular and submolecular levels, combining two microscopy techniques. However, the main limitation of correlative microscopy is to develop a labeling technique that can be easily used first in light and then in electron microscopy. Laser etching is a well-established method to create precisely designed shapes or volumes in various materials including glass. We have applied this technique to develop a new correlative light and electron microscopy method and to apply it in our study of the Golgi apparatus. The location of the cell of interest is laser-inscribed into the glass allowing a simple follow-up in light and fluorescence microscopy. Furthermore, the glass surface is laser-etched and upon fixation and flat embedding, the inverse ridge can be localized as well as the cell of interest, which is then processed for electron microscopy.
Colombelli
Julien
J
Cell Biology and Cell Biophysics Unit, EMBL, Heidelberg, Germany.
Tängemo
Carolina
C
Haselman
Uta
U
Antony
Claude
C
Stelzer
Ernst H K
EH
Pepperkok
Rainer
R
Reynaud
Emmanuel G
EG
D002460:Cell Line; D006056:Golgi Apparatus / Q000648:ultrastructure; D007834:Lasers; D008854:Microscopy, Electron / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D016613:Plastic Embedding; D013194:Staining and Labeling