Cryo-CLEM inside the Aquilos Cryo-FIB using a new integrated fluorescence light microscope
Alexander Rigort
Thermo Fisher Scientific, FEI Deutschland GmbH, Fraunhofer Strasse 11, 82152 Martinsried, Germany
Abstract
Cellular cryo-electron tomography is a high-resolution technique that enables imaging of the molecular machinery of a cell at close-to-native conditions. To render cells suitable for tomographic imaging, electron-transparent regions, named cryo-lamellae, must first be prepared with a cryo-focused ion beam (cryo-FIB) microscope. These cryo-lamellae are essentially snapshots of functional cellular environments preserved in a fully hydrated state and embedded in non-crystalline vitreous ice. To enable correlative imaging of vitrified samples, a new integrated light microscope has been added to the cryo-FIB microscope. With the iFLM Correlative System, the correlation of signals between two imaging modalities - light and electron microscopy - within one instrument becomes possible. The built-in fluorescence module facilitates localization of lamella sites and makes it possible to verify whether already milled lamellae contain fluorescent targets.