Literature DB >> 24106925

Controlling FIB-SBEM slice thickness by monitoring the transmitted ion beam.

K M Boergens1, W Denk.   

Abstract

Serial block-face electron microscopy with focused ion beam cutting suffers from cutting artefacts caused by changes in the relative position of beam and sample, which are, for example, inevitable when reconditioning the ion gun. The latter has to be done periodically, which limits the continuous stack-acquisition time to several days. Here, we describe a method for controlling the ion-beam position that is based on detecting that part of the ion beam that passes the sample (transmitted beam). We find that the transmitted-beam current decreases monotonically as the beam approaches the sample and can be used to determine the relative position of beam and sample to an accuracy of around one nanometre. By controlling the beam approach using this current as the feedback parameter, it is possible to ion-mill consecutive 5 nm slices without detectable variations in thickness even in the presence of substantial temperature fluctuations and to restart the acquisition of a stack seamlessly. In addition, the use of a silicon junction detector instead of the in-column detector is explored.
© 2013 The Authors Journal of Microscopy © 2013 Royal Microscopical Society.

Entities:  

Keywords:  Beam position control; FIB-SBEM; FIB-SEM; focused ion beam; slice thickness

Year:  2013        PMID: 24106925     DOI: 10.1111/jmi.12086

Source DB:  PubMed          Journal:  J Microsc        ISSN: 0022-2720            Impact factor:   1.758


  6 in total

1.  Simultaneous determination of sample thickness, tilt, and electron mean free path using tomographic tilt images based on Beer-Lambert law.

Authors:  Rui Yan; Thomas J Edwards; Logan M Pankratz; Richard J Kuhn; Jason K Lanman; Jun Liu; Wen Jiang
Journal:  J Struct Biol       Date:  2015-10-09       Impact factor: 2.867

Review 2.  Challenges of microtome-based serial block-face scanning electron microscopy in neuroscience.

Authors:  A A Wanner; M A Kirschmann; C Genoud
Journal:  J Microsc       Date:  2015-04-23       Impact factor: 1.758

3.  Enhanced FIB-SEM systems for large-volume 3D imaging.

Authors:  C Shan Xu; Kenneth J Hayworth; Zhiyuan Lu; Patricia Grob; Ahmed M Hassan; José G García-Cerdán; Krishna K Niyogi; Eva Nogales; Richard J Weinberg; Harald F Hess
Journal:  Elife       Date:  2017-05-13       Impact factor: 8.140

4.  Image-based correction of continuous and discontinuous non-planar axial distortion in serial section microscopy.

Authors:  Philipp Hanslovsky; John A Bogovic; Stephan Saalfeld
Journal:  Bioinformatics       Date:  2017-05-01       Impact factor: 6.937

Review 5.  Microscopic Visualization of Cell-Cell Adhesion Complexes at Micro and Nanoscale.

Authors:  Bieke Vanslembrouck; Jian-Hua Chen; Carolyn Larabell; Jolanda van Hengel
Journal:  Front Cell Dev Biol       Date:  2022-04-20

6.  From Light Microscopy to Analytical Scanning Electron Microscopy (SEM) and Focused Ion Beam (FIB)/SEM in Biology: Fixed Coordinates, Flat Embedding, Absolute References.

Authors:  Manja Luckner; Gerhard Wanner
Journal:  Microsc Microanal       Date:  2018-09-24       Impact factor: 4.127

  6 in total

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