Research Project
8590991
DESCRIPTION (provided by applicant): Project Summary / Abstract Engineering Arts (EA) proposes the development of a detachable nozzle piezoelectric dispense system for use in Cryo-TEM (Cold - Transmission Electron Microscopy) sample preparation instrumentation. Cryo-TEM is an important tool for studying the structure of biological macromolecules in their native aqueous state. The challenging and laborious process of preparing 'thin-ice' frozen aqueous samples for Cryo-TEM has remained essentially unchanged for decades. Typically a single aqueous biological sample (~3¿l) is manually applied onto a thin porous Electron Microscopy (EM) grid which is then blotted to remove excess sample and then plunged into liquid-ethane for flash freezing. This process hopefully leaves enough regions on the grid with the proper vitrified 'thin-ice', typically 50 to 200nm thickness, required for EM imaging. The detachable piezoelectric nozzle dispense system proposed here is a key innovation that will lead to productivity, quality and capability breakthroughs for Cryo-TEM workflow. The following key benifits for Cryo-TEM process will be realized: 1. Apply many different samples (or sample conditions) to each Cryo-TEM grid. 2. Increase process control and yields of acceptable thin-ice over current standard blotting methods. 3. Lower starting sample volume requirements and consumption per grid. 4. Allow time-resolved study of mixing / binding events of multiple samples. EA will design, fabricate and optimize a multichannel detachable nozzle piezoelectric dispense system that is capable of dispensing droplets (less than 50 picoliters) of several different biological samples (or sample conditions) onto each TEM grid. A three channel detachable nozzle piezoelectric dispense system retrofitted onto a custom Cryo-TEM sample preparation instrument (Spotiton instrument platform) at NRAMM (National Resource for Automated Molecular Microscopy, Scripps, La Jolla CA.) for testing, optimization and validation.
