A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:05Title
- 01:11Principle of the Random Conical Tilt Method
- 02:10Preparation of Holey Carbon Grids Covered with Thin Carbon
- 05:08Negative Staining of the Exosome Complex
- 06:23Electron Microscopy of the Exosome Complex
- 08:10Image Processing of the Data
- 10:55Representative Results for Single Particle Electron Microscopy Reconstruction of the Exosome Complex Using the Random Conical Tilt Method
- 11:29Conclusion
Questo articolo descrive un metodo standard per ottenere un modello tridimensionale (3D) ricostruzione delle macromolecole biologiche mediante microscopia elettronica a colorazione negativa (EM). In questo protocollo, viene spiegato come ottenere la struttura 3D del complesso exosome Saccharomyces cerevisiae a media risoluzione con il metodo casuale conica ricostruzione tilt (RCT).
