Acquisition of a cryogenic holder for cryo-electron microscopy

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Cells have elaborate systems for organizing their internal spaces, including the ability to form specialized compartments and skeleton-like structures that ensure proper positioning of cellular components. Maintaining the correct internal cellular organization is critically important in all forms of life. My research is aimed at understanding complex organization in bacterial cells. We do this by studying the structures of the tiny molecular machines that drive these organizing processes to give us insight into how they function. One of the most powerful tools at our disposal is the ability to examine molecular complexes in three dimensions using modern electron microscopes. In order to achieve three-dimensional structures samples must be rapidly frozen, which preserves their fine structure. The challenge we then face is maintaining the samples at extremely low temperatures during transfer into the microscope and during imaging. This requires specialized sample holders, which can be maintained near the temperature of liquid nitrogen (-196 degrees C), while remaining stable enough to not drift while we acquire images of our samples at high magnification. This proposal requests a state-of-the-art cryo holder, which will allow us to visualize individual molecular structures at high resolution in three dimensions.