Oct 31, 2018

Oxygen plasma focused ion beam scanning electron microscopy for biological samples

BioRxiv : the Preprint Server for Biology
Sergey GorelickAlex de Marco

Abstract

Over the past decade, gallium Focused Ion Beam-Scanning Electron Microscopy (FIB-SEM) has been established as a key technology for cellular tomography. The utility of this approach, however, is severely limited both by throughput and the limited selection of compatible sample preparation protocols. Here, we address these limitations and present oxygen plasma FIB (O-PFIB) as a new and versatile tool for cellular FIB-SEM tomography. Oxygen displays superior resin compatibility to other ion beams and produces curtain-free surfaces with minimal polishing. Our novel approach permits more flexible sample preparation and 30% faster data collection when compared to using gallium ion sources. We demonstrate this alternative FIB is applicable to a variety of embedding procedures and biological samples including brain tissue and whole organisms. Finally, we demonstrate the use of O-PFIB to produce targeted FIB-SEM tomograms through fiducial free en-block correlative light and electron microscopy.

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Mentioned in this Paper

FIB-P Protocol
SHFM1
X-Ray Computed Tomography
Ions
Embedding
Electron Microscopy, Diagnostic
AS resin
Electron Microscopy
Plasma
Tomography

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