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Biochemistry
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微图案透射电子显微镜网格,用于在全细胞冷冻电子断层扫描工作流程中指导细胞定位
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal
Biochemistry
Micropatterning Transmission Electron Microscopy Grids to Direct Cell Positioning within Whole-Cell Cryo-Electron Tomography Workflows
Please note that all translations are automatically generated.
Click here for the English version.
微图案透射电子显微镜网格,用于在全细胞冷冻电子断层扫描工作流程中指导细胞定位
DOI:
10.3791/62992-v
•
09:53 min
•
September 13, 2021
•
Bryan S. Sibert*
1,2,3
,
Joseph Y. Kim*
1,4
,
Jie E. Yang
2,3
,
Elizabeth R. Wright
2,3,5
1
Department of Biochemistry
,
University of Wisconsin, Madison
,
2
Cryo-Electron Microscopy Research Center, Department of Biochemistry
,
University of Wisconsin, Madison
,
3
Midwest Center for Cryo-Electron Tomography, Department of Biochemistry
,
University of Wisconsin, Madison
,
4
Department of Chemistry
,
University of Wisconsin, Madison
,
5
Morgridge Institute for Research
Chapters
00:04
Introduction
01:25
Preparation of Grids for Patterning and Application of the Anti-fouling Layer
02:50
Applying PLPP Gel
03:33
Micropatterning
06:11
Deposition of Extracellular Matrix (ECM) Proteins
06:54
Results: Micropatterning to Direct Cell Positioning within Whole-cell Cryo-electron Tomography
09:17
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
该协议的目标是将细胞粘附和生长引导到冷冻电子显微镜的网格目标区域。这是通过应用抗污垢层来实现的,该抗污垢层以用户指定的模式消融,然后在细胞接种之前在图案化区域沉积细胞外基质蛋白。
Tags
Micropatterning
Transmission Electron Microscopy
Cryo-electron Tomography
Cell Positioning
Whole-cell Cryo-electron Tomography
EM Grids
Poly-L-lysine
PEG-SVA
HEPES
Cell Culture
Cryo-CLEM
Cryo-FIB Milling
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