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The Peel-Blot Technique: A Cryo-EM Sample Preparation Method to Separate Single Layers From Multi-Layered or Concentrated Biological Samples
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生物化学
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JoVE 杂志 生物化学
The Peel-Blot Technique: A Cryo-EM Sample Preparation Method to Separate Single Layers From Multi-Layered or Concentrated Biological Samples

The Peel-Blot Technique: A Cryo-EM Sample Preparation Method to Separate Single Layers From Multi-Layered or Concentrated Biological Samples

DOI:
10.3791/64099-v

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07:27 min

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June 29, 2022

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, ,
1School of Biological Sciences,Georgia Institute of Technology, 2Department of Structural Biology,Genentech, 3School of Chemistry and Biochemistry,Georgia Institute of Technology

Chapters

  • 00:04Introduction
  • 00:33Preparation Steps for the Peel‐Blot Technique
  • 01:57Placing the Carbon Film onto the Grid
  • 03:07Separation of Multi-Layered 2D Crystals into Single Layer
  • 04:55Results: 2D Crystals of Human Leukotriene C4 Synthase and Liposomes Subjected to the Peel‐Blot Method, the Peel-Blot Footprint, and Blotting an EM Grid Prepared by Back-Injection on a Submicron Filter Paper
  • 06:42Conclusion

Summary

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The peel-blot technique is a cryo-EM grid preparation method that allows for the separation of multilayered and concentrated biological samples into single layers to reduce thickness, increase sample concentration, and facilitate image processing.

Tags

Keywords: Peel-Blot TechniqueCryo-EMSample PreparationMulti-Layered SamplesConcentrated SamplesSingle LayersThickness ReductionConcentration IncreaseImage ProcessingFilter PaperParaffin FilmSubmicron Filter PaperAnti-Capillary Forceps600 Mesh GridTrehalose SolutionMicaCarbon FilmGrid Orientation

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