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Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy
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Biologia
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JoVE Journal Biologia
Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy

Targeted Studies Using Serial Block Face and Focused Ion Beam Scan Electron Microscopy

DOI:
10.3791/59480-v

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09:09 min

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August 10, 2019

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, , ,
1VIB Bio Imaging Core, 2VIB Inflammation Research Center, 3Department of Molecular Biomedical Research,UGent

Capitoli

  • 00:04Titolo
  • 00:51Sample Fixation and Processing for Electron Microscopy
  • 03:51Prepare Embedded Samples for Imaging
  • 05:03Imaging in the SBF-SEM (Serial Block Face Scanning Electron Microscopy) and Data Processing
  • 06:22Imaging in the FIB-SEM (Focused Ion Beam SEM)
  • 07:49Results: SBF-SEM and FIB-SEM Data
  • 08:46Conclusion

Summary

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Here, we present a protocol for efficiently combining serial block face and focused ion beam scanning electron microscopy to target an area of interest. This allows for efficient searching, in three dimensions, and locating rare events in a large field of view.

Tags

Serial Block Face Scanning Electron Microscopy (SBF-SEM)Focused Ion Beam Scanning Electron Microscopy (FIB-SEM)3D UltrastructureArabidopsis ThalianaRoot TipsSample PreparationFixationPost-fixationOsmium TetroxideRuthenium RedThiocarbohydrazide (TCH)DehydrationMicrowavePhosphate Buffer

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