Site-Specific Cryo-focused Ion Beam Sample Preparation Guided by 3D Correlative Microscopy.

ABSTRACT: The development of cryo-focused ion beam (cryo-FIB) for the thinning of frozen-hydrated biological specimens enabled cryo-electron tomography (cryo-ET) analysis in unperturbed cells and tissues. However, the volume represented within a typical FIB lamella constitutes a small fraction of the biological specimen. Retaining low-abundance and dynamic subcellular structures or macromolecular assemblies within such limited volumes requires precise targeting of the FIB milling process. In this study, we present the development of a cryo-stage allowing for spinning-disk confocal light microscopy at cryogenic temperatures and describe the incorporation of the new hardware into existing workflows for cellular sample preparation by cryo-FIB. Introduction of fiducial markers and subsequent computation

SUBMITTER: Arnold J 

PROVIDER: S-EPMC4775854 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature