3D quantification of a fluorescent marker in rodent tissues and organs

Your Needs:
  • Visualize anatomical structures in 3D
  • Localize and quantify a specific fluorescent marker in 3D
  • Preclinical study of treatment efficacy

Our Solutions:
  • Light sheet microscopy after sample clearing
  • Robust procedure to image an entire tissue or organ at different scales
  • Semi-automated process to characterize structures and localize markers in 3D

General Procedure

Prior to sample collection by Imactiv-3D:
  • Sample optical clearing
  • In vivo labeling by infusion with a fluorescent marker of interest before euthanasia
  • Formalin fixation of extracted sample

Image acquisition:
  • Sample clearing
  • 3D light sheet fluorescence microscopy with multi-position acquisition

Image processing and analysis:
  • Segmentation of anatomical structures based on auto-fluorescence information (in grey)
  • Quantification of fluorescence intensity within the identified volumetric structures (in red)

Application example

  • Aim:quantification of a fluorescent marker in a mouse leg with a tumor compared to a healthy one.
  • The auto-fluorescence signal intensity was used for the 3D identification of different anatomical structures (cortical bone, epiphysis, tumor), independently of the marker fluorescence. Their volumes were reconstructed.
  • The marker fluorescence intensity was measured within the identified volumetric structures and compared between healthy leg and tumoral leg.