3D ex vivo imaging and characterization of cell death

Your Needs:
  • Study of cell death in any organ
  • Preclinical study of treatment efficacy

Our Solutions:
  • Light sheet microscopy after clearing of ex vivo samples
  • Automated 3D image processing to count necrotic cells in organs

General Procedure

Prior to sample collection by Imactiv-3D:
  • In vivo animal perfusion to label necrotic nuclei (PI)
  • Formalin fixation of extracted sample

Image acquisition:
  • Staining of ex vivo samples with nuclear dye to label all cells (DRAQ5)
  • Biopsies of 2.5-mm-diameter regions of interest
  • Clearing of samples and 3D light sheet microscopy

Image processing and joint analysis of both fluorescence channels:
  • 3D image restoration based on denoising and pre-segmentation
  • 3D refined segmentation and quantification
  • Quantitative analysis of necrosis rate as the ratio: number of necrotic (PI) nucleitotal number (DRAQ5) of nuclei

Application example: Quantification of necrosis after myocardial infarction

  • Definition of regions of interest depending on their distance to the infarct scar. In this study, 21 regions were explored (triplicate in each region).
  • Analysis of necrotic nuclei percentage in healthy and post-infarct rat hearts.
  • Regions near the scar showed much more necrotic cells (regions 1 to 3) than the control region (region 4) and than all regions in the healthy heart.