Scanning Transmission Electron Tomography of Blood Platelets in Thick Sections

Abstract

bearing nitrilotriacetic acid substituents at the end of a poly(ethylene glycol)grafted surface that promotes specific capture of protein targets for single particle reconstruction analysis. The utilization of these grids for specific adsorption of the targeted protein onto the grid surface results in wellcontrolled surface concentration enhancements and a days-to-minutes reduction in time required for the preparation of a purified sample for cryoEM analysis from an E. coli expression system. The selective and reversible capture of his-tag T7 bacteriophage and GroEL from crude lysates, as well as purified nanodisc-solubilized his-malFGK2, on these NTA-modified grids with an exceptionally low level of adsorption by non-target proteins has been observed. Our data illustrates the utility of these grids for selective capture from complex mixtures, detergent-solubilized membrane protein isolates, and expression systems yielding low copy numbers of the desired target in a manner that is well-suited for single particle reconstruction analysis.