Q:

How can I image cells in a RAFT™ 3D Cell Culture?

A:
Below is a list of compatible imaging techniques and systems with RAFT™ Cultures, depending on the type of cells being cultured. RAFT™ Culture Types: Cells that tend to maintain a round morphology or that form higher order structures such as tumouroids (e.g. cancer cells such as MCF7, HCT116), Cells that tend to elongate such as fibroblasts (e.g. primary human dermal fibroblasts) Live imaging techniques: Brightfield, Phase contrast (may be an issue as the higher concentration collagen matrix stops some light), Non-toxic fluorescent dyes (e.g. Promega’s CellTox Green) End point analysis techniques (where you are prepared to kill the cells): Immunofluorescence, Fluorescent dyes Protocol changes compared to 2D: So far, little or no changes from 2D protocols have been needed with RAFT™ cultures. Imaging systems compatible: Widefield microscopes (brightfield and fluorescent), Confocal microscopes, An automated z-stage can be helpful if trying to image the whole thickness of the RAFT™ culture, High content analyzers have also been used successfully to study RAFT™ 3D cultures, such as the Acumen (TTP Labtech) or Opera (Perkin Elmer)
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Categories:
3D Cell Culture
Research Areas:
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