HIGH-CONTENT IMAGING
Cell counting
Which cell populations respond to a compound?
A total cell count can hide differences between populations. High-content microscopy separates labelled populations within the same culture and measures how their cell numbers change with treatment.
EXPERIMENTAL DATA
The method
Treat
Expose mono-cultures or co-cultures to a concentration series alongside vehicle and reference controls.
Identify
Label nuclei and, where required, use population-specific markers or live/dead stains. Acquire images with automated microscopy.
Quantify
Segment cells, assign marker-defined populations and compare counts across conditions. Normalise concentration-response profiles to the relevant controls.
Reading the results
Total and population-specific cell counts, relative responses and concentration-response curves. Viability readouts can be added with an appropriate live/dead stain.
Interpretation
Fewer cells can reflect reduced proliferation, cell death or both. Cell counts alone do not distinguish these mechanisms; complementary proliferation and cell-death measurements help resolve them.
Study design
Cell model, population markers, treatment duration and image-analysis settings are selected for the study. The example illustrates a measured response in one co-culture, not selectivity across all biological models.