Aim: Slide-based cytometry (SBC) allows to "ask a cell a second time." We used this tool for detailed immunophenotyping of peripheral blood leukocytes (PBLs).
Methods: PBLs primarily stained for CD-markers and DNA were immobilized on a glass slide and analyzed by laser scanning cytometry. Then, iterative restaining was applied for a second and a third analysis. Based on the cells' fixed location, analyses were merged on a single-cell level.
Results: We analyzed six virtual immunostainings by "recycling" the PE-channel for four CD-markers.
Conclusion: Iterative restaining might prove to be a pivotal tool for n-color immunophenotyping exclusive to SBC concepts.
(c) 2006 International Society for Analytical Cytology.