Iterative restaining as a pivotal tool for n‐color immunophenotyping by slide‐based cytometry

Abstract
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. © 2006 International Society for Analytical Cytology