Application Note: Label-Free, Real-Time Live-Cell Assays for 3D Organoids Embedded in Matrigel®

Organoid technologies are increasingly being used as in-vitro models of human development and disease. This is because they exhibit structural, morphogenetic and functional properties that recapitulate in-vivo pathophysiology. To successfully use these models across a variety of research disciplines and applications, approaches that reduce variability and technology pipelines to image and quantify these complex cell models are required.

Currently, techniques to robustly characterize and visualize these models, may be limited by some approaches, including a requirement to label cells (fluorescence-based quantification).  This may be challenging and not amenable to a range of cell types.

In this application note, we demonstrate the use of the Incucyte® Live-Cell Analysis System, in combination with the Incucyte® Organoid Analysis Software Module, to simplify and facilitate temporal assessment of organoid growth or death, through integrated, real-time label-free metrics. Automated software continuously locates and analyzes embedded organoids in physiologically relevant conditions.

Included Highlights:

  • The ability to kinetically visualize and quantify distinct organoid morphologies, and segment individual objects embedded in Matrigel® across a range of cell types is demonstrated.
  • An example of this approach to visualize and quantify cystic fibrosis transmembrane conductance regulator (CFTR) function, thereby enabling label-free assessment of cystic fibrosis in-vitro, is provided.
  • The use of integrated, real-time label-free metrics, to assess drug-induced cellular changes and characterize mechanisms of action in a 96-well plate format, is shown.

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