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

Project Summary 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 researches 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 la rge 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 all ow 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. Project Narrative: This proposal will develop tools that allow biologists to get a clearer picture of the structure and function of the cell. This knowledge is funda mental to innumerable areas of medical research and practice.