2006•Cytometry Part ARequires access

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

Wiebke Laffers, Anja Mittag, Dominik Lenz, Attila Tárnok, Andreas O. H. Gerstner

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.

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Key concepts: Immunophenotyping, Cytometry, Flow cytometry, Peripheral blood, Pathology, Computer science, Medicine, Immunology