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

Chapitres

  • 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

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Traduction automatique

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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