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Technical Note

3D Volumetric Analysis of Luminal Spaces Inside Cysts or Organoids

Introduction

High-content assays using 3D objects such as cysts or organoids can be challenging from the perspectives of both image acquisition and image analysis. In this technical note we describe how to image and analyze epithelial Madin-Darby canine kidney (MDCK) cysts in 3D on the Operetta CLS high-content analysis system.

We address:

  • How to reduce the loss of image resolution as you image deeper into the sample
  • How to enable 3D segmentation and analysis for your 3D high-content assays
  • How to increase throughput for 3D imaging through shorter exposure times
  • How to eliminate long and tedious data transfer steps