Culture spheroids – Cancer cells can be cultured directly in an ultra-low attachment (ULA), round bottom plate, or other labware to develop the typical morphology of a spheroid. Other labware allows one to grow multiple spheroids in a single well.
Treat with compounds – After spheroid formation, compounds at the desired concentrations are added into the wells, and then incubated for one to several days, depending on the mechanism being studied.
Optional Step: Check cell viability
Stain for markers – After compound treatment is complete, stains are added directly to the media. Stains that require no washing can be used to avoid disturbing spheroids, but spheroids can be carefully washed, even using automation, if necessary.
Acquire spheroid images – Images within the body of the spheroid can be captured individually or as a z-stack (multiple images taken at differing depths) using specialized imaging equipment.
Analyze cancer cells – Use cellular imaging analysis software to run quantitative analysis of the cell images to monitor the expression of different markers and to quantify biological readouts.
Hover over a step above for additional information.
ImageXpress Micro Confocal High-Content Imaging System
High-Content Image Acquisition and Analysis Software
SpectraMax i3x Multi-mode Microplate Reader