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Biochemistry

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Preparation of Sample Support Films in Transmission Electron Microscopy using a Support Floatation Block

4.4K Views

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06:10 min

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April 8th, 2021

DOI :

10.3791/62321-v

April 8th, 2021

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Natàlia de Martín Garrido¹, Kailash Ramlaul¹, Christopher H. S. Aylett¹

¹Section of Structural & Synthetic Biology, Department of Infectious Diseases, Faculty of Medicine, Imperial College London, South Kensington Campus

Transcript

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Summary

Sample preparation for cryo-electron microscopy (cryo-EM) is a significant bottleneck in the structure determination workflow of this method. Here, we provide detailed methods for using an easy-to-use, three-dimensionally printed block for the preparation of support films to stabilize samples for transmission EM studies.

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Transmission Electron Microscopy

Cryo EM Samples

Flotation Block

Sample Preparation

Hydrophilic Treatment

Bolting Devices

Negative Staining

Graphene Oxide Suspension

Air Water Interface

Safety Precautions

Chapters in this video

0:04

Introduction

0:47

Preparation of Negatively Stained Samples with Amorphous Carbon Support

2:21

Preparation of Graphene Oxide-coated TEM Grids

3:27

Preparation of Samples on Monolayer-graphene Films Using the Flotation Block

4:44

Results: Examples of Sample Support Films Prepared Using the Floatation Block

5:34

Conclusion

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