Speaker
Description
In-situ cryo electron tomography (cryo-ET) is a powerful technique to visualize cellular components in native context and near-atomic resolution. The workflow often involves the use of a focused ion beam (FIB) to thin down the vitrified cell into a ~200 nm thick “cryo lamella”. Considering the low abundance of certain subcellular structures and the extremely limited volume of the cryo lamella, two of the most important challenges in the workflow are as follows: 1) Localizing the cells that contain the regions of interest (ROIs) (2D localization)
2) Capturing the ROIs in the limited volume of the lamella during FIB milling (3D localization) To address these challenges, we present a high-quality integrated fluorescent microscope, called METEOR. Together with its dedicated and user-friendly software ODEMIS, METEOR provides fast and accurate 2D correlation between fluorescence (FLM) and scanning electron microscopy (SEM) images resulting in efficient 2D localization of the ROIs. Moreover, the high-resolution METEOR z-stacks can effectively be used for precise 3D correlation between FLM and FIB images, resulting in an optimized and targeted FIB milling process with high throughput.
| Authors | Deniz Daviran, Kevin Homberg*, Marit Smeets |
|---|---|
| Keywords | Cryo-ET, Cryo-FIB, CLEM |
