More Content from High-Content Screening: Analyzing Specimen in 3D
Alexander Schreiner,
Molecular Cell Biologist,
PerkinElmer
When developing a screening assay one of the most critical aspects is the choice of cellular model including the decision to choose between 2D or 3D approaches. Multicellular “oids” (tumoroids, spheroids, organoids) bear the potential to better predict drug candidate effects during preclinical screening. However, 3D models require more effort and compared to 2D cultures, 3-dimensional assays tend to be more complex across all steps of the workflow. Reliably generating sufficiently large numbers of uniform spheroids for screening and imaging of 3D volumes at high quality are among the challenges. Using 3D spheroids and cysts as examples, we will show experimental workflows, explaining how to effectively grow these models using ULA coated U-bottom plates or low concentration gels. The careful selection of dyes and clearing strategies can improve the image quality while targeted imaging of spheroids helps to significantly shorten imaging times and to minimize the data volume. Nevertheless, being able to effectively analyze spheroids is still one of the main bottlenecks. Dedicated high-content software tools can help users to, speed up 3D image acquisition by targeted imaging of objects of interest, better understand the spatial context of your 3D cell models through different visualization methods, measure volume and morphology changes in 3D to characterize your specimen in detail and export 3D movies to share and publish your results.
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