Single Particle Electron Microscopy Reconstruction of the Exosome Complex Using the Random Conical Tilt Method

The overall goal of the following experiment is to get the three dimensional structure of the exosome complex using the random conical tilt method. This is achieved by first depositing a thin layer of carbon film onto the holy carbon grid to make the support substrate for the protein complex and the stain as a second step. The specimen and the stain are applied onto the grid, which buries the specimen into heavy metal salts.

Electron microscopy is then carried out to get tilt pairs of micrographs of the specimen. Finally, results are obtained that show the three dimensional structure of the exosome complex based on image analysis using the random conical tilt method. Generally, individuals new to this method will struggle because sometimes good preparation of specimen for random conical tilt method is difficult as well as the procedure for the image processing that follows View.