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Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids
JoVE Journal
Biologie
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JoVE Journal Biologie
Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids

Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids

DOI:
10.3791/60709-v

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10:40 min

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June 05, 2020

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, , , ,
1Princess Máxima Center for Pediatric Oncology, 2Department of Cancer Research, Oncode Institute,Hubrecht Institute-KNAW Utrecht, 3Cancer Genomics Center (CGC), 4Hubrecht Institute,Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center (UMC) Utrecht

Kapitel

  • 00:04Introduction
  • 00:46Organoid Recovery
  • 02:28Fixation and Blocking
  • 03:30Immunolabeling
  • 05:33Optical Clearing
  • 06:08Confocal Imaging Slide Preparation
  • 07:01Image Acquisition and Processing
  • 08:06Results: Representative 3D Imaging and Organoid Clearing
  • 10:01Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

The entire 3D structure and cellular content of organoids, as well as their phenotypic resemblance to the original tissue can be captured using the single-cell resolution 3D imaging protocol described here. This protocol can be applied to a wide range of organoids varying in origin, size and shape.

Tags

Organoids3D ImagingSingle-cell ResolutionOptical ClearingSample PreparationBasement Membrane ExtractRecovery SolutionPBS-BSACentrifugationParaformaldehydePBS-Tween 20Organoid Washing Buffer

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