MRI: Acquisition of Integrated Instrumentation to Facilitate Correlative Light and Electron Microscopy of Cellular Systems

Brandeis University, Waltham MA

Investigators

Abstract

This award is for the acquisition of instrumentation needed to establish a facility that will provide a new window into the functional organization of cells. The facility will contain an integrated system of instruments for cryo-preparation to facilitate two research areas: 1) correlative light and electron microscopy (CLEM), and 2) cryo-electron tomography of vitreous sections. Cells have complex interactions among intracellular components, each of which exhibits precise spatial localization and temporal dynamics. Advances in fluorescence light microscopy allow precise tracking of individually labeled molecules; this can provide important insights into dynamic cellular processes of living cells with high temporal resolution. However, current light microscope technology lacks the resolution of EM to reveal structural details at a molecular level with high spatial resolution. EM images provide only static snapshots of dynamic intracellular processes. The instrumentation acquired through this award will enable a combination of the advantages of light and EM in two ways: 1) Tracking the dynamics of individual intracellular structures in a living cell by time-lapse imaging, followed by rapid freezing of the specimen and subsequent analysis of the same cell by EM and three-dimensional (3D) reconstruction methods; 2) Scanning large regions of cells or tissues to identify specific sites for EM analysis that occur at low frequency, enabling EM studies of cellular structures that would not otherwise be feasible. This correlative approach will provide new insights into: 1) the structure and function of macromolecules, cells, and tissues, and 2) the dynamic spatiotemporal intra- and intercellular interactions that regulate cellular function. This new facility will train a new generation of scientists who will apply correlative light and electron microscopy to carry out innovative experiments in cell biology and neurobiology. The facility will be unique in the greater Boston area and will facilitate interactions and collaborations among investigators at the university and other institutions in the area. The facility will train undergraduates, graduate students and postdoctoral fellows in cutting-edge technologies. New theoretical and practical courses in imaging techniques and applications will be developed and implemented; these new courses will be open to members of neighboring institutions. The university hosts a summer research internship program for minority undergraduates from other campuses. Interns from this program will have access to this facility as part of their research program. Over 40% of the Life Sciences faculty are female, and this award is spearheaded by three women faculty, providing excellent role models for students. This facility will integrate education, training and research to ultimately provide new fundamental insights into cellular architecture and function.