System for generation of frozen-hydrated samples using the Focused Ion Beam

DESCRIPTION (provided by applicant): The long term objective of our application is to develop hardware and procedures that will allow researchers to better understand the function of the cell. Our specific aim is to provide a system that will allow researchers to better prepare frozen hydrated specimens for study via tomographic reconstruction in the transmission electron microscope(TEM) Current methods of preparing frozen hydrated specimens using cryo-ultramicrotomy produce wavy samples with tissue deformation and require a large investment of time and patience to get even low yield results. A commercial focused ion beam (FIB) system is a promising alternative tool for sample preparation, but the latest research suggests that currently available systems for use with the FIB allow sample contamination (primarily frost) and have poor sample handling hardware for this technique. In close collaboration with the Resource for the Visualization of Biological Complexity (RVBC) at the Wadsworth Center's NIH Biotechnological Resource, we will develop a new sample holder, demonstrate smooth transfer of this sample into and out of the FIB, demonstrate basic FIB cutting patterns and ultimately show superior tomograms. It is our intent to develop and commercialize a cost effective system that will enable these techniques to become widely available to the biology research community. Public comment: This proposal will develop tools that allow biologist to obtain images of the structure and function of the cell at unprecedented levels of resolution and thus clarity. This knowledge is fundamental to our basic knowledge of biological function, and will directly impact medical research and practice.