Cryogenic Electron Tomography (cryoET) is an emerging technique that can resolve subcellular structures in situ with the potential to reach near atomic resolutions.  Correlated cryo-fluorescence light microscopy (cryoFLM) and cryoET of frozen, hydrated cells can be used to study cellular and molecular functions and dynamics in the 3D context of cells and tissues at a higher resolution than any other imaging technique. For samples which are too thick for imaging via cryoET, a focused ion beam (FIB) can be used to produce thin lamellae (100-300 nm) from vitrified cells or tissues containing targets of interest. The embedded cryoFLM in the cryoFIB Scanning Electron Microscope (cryoFIB-SEM) can guide the search of the targets to form the lamellae which are then imaged by cryoET in a cryogenic transmission electron microscope. 

Additionally, cryo-volume Electron Microscopy (cryo-vEM) can be performed to produce 3D image stacks from serially FIB-milled and SEM imaged surface areas specimens ranging from unicellular algae to multicellular organisms and tissue slices.  The SCSC Helios Hydra Bio Plasma FIB-SEM (Hydra Bio PFIB) with high resolution optics and imaging detectors is ideally suited for cryo-vEM applications, using different gas species for FIB-milling.  Generation of a lamella at a targeted region of interest within the cryo-vEM workflow, consequently integrates cryo-vEM with cryoET.

MISSION STATEMENT:

The mission of the Stanford-SLAC CryoET Specimen Preparation Center (SCSC) is to provide:

• Access to state-of-the-art instrumentation and knowledge about cryoET specimen preparation
• Training and support, to increase the ability of the cryoET community to perform experiments particularly in Life and Biomedical Sciences
• Development of workflows and implementation of new technologies for cryoET specimen preparation

The Center implements state-of-the-art cryo-specimen preparation devices. We provide expert staff that train, assist, and advise users both onsite and remotely. We cross-train scientists who want to employ cryoET within their own research portfolios. Our training is targeted at a wide variety of skill levels - including short to long-term in-residence training programs for cryoET specimen preparation. Access to the Center is through an open process based on scientific merit. Specimens prepared at SCSC can be transferred for data collection at the investigator's lab or at the Center Hub in Madison, Wisconsin.

Read about SCSC