An improved method for discrimination of cell populations in tissue sections using microscopy-based multicolor tissue cytometry.

Background: In tissue context, researchers and pathologists lack a generally applicable standard for quantitative determination of cytological parameters. Increasing knowledge of disease-specific markers calls for an appropriate in situ tissue cytometry.

Methods: Microscopy-based multicolor tissue cytometry (MMTC) permits multicolor analysis of single cells within tissue context.

Results: Tissue specimens stained for CD45/CD3/CD4/CD8 were analyzed. Specificity as well as reproducibility of MMTC is demonstrated and a novel MMTC-based function to improve visual discrimination of subpopulations is introduced.

Conclusions: Our data demonstrate that MMTC constitutes an important step toward automated and quantitative fluorometry of solid tissues and cell monolayers.