Iterative restaining as a pivotal tool for n-color immunophenotyping by slide-based cytometry.
OBJECTIVE
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.


CONCLUSIONS
Iterative restaining might prove to be a pivotal tool for n-color immunophenotyping exclusive to SBC concepts.
Department of Otorhinolaryngology/Surgery, University of Bonn, D-53105 Bonn, Germany.
Wiebke
W
Laffers
Anja
A
Mittag
Dominik
D
Lenz
Attila
A
Tárnok
Andreas O H
AO
Gerstner
D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D018941:Antigens, CD19 / Q000032:analysis; D017252:CD3 Complex / Q000032:analysis; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens / Q000032:analysis; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007091:Image Processing, Computer-Assisted; D016130:Immunophenotyping / Q000379:methods*; D047569:Laser Scanning Cytometry / Q000379:methods*; D017493:Leukocyte Common Antigens / Q000032:analysis; D007962:Leukocytes / Q000737:chemistry / Q000166:cytology / Q000276:immunology; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D016131:Lymphocyte Subsets / Q000737:chemistry / Q000166:cytology; D013194:Staining and Labeling / Q000379:methods*
D000906:Antibodies; D018941:Antigens, CD19; D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D005456:Fluorescent Dyes; D018950:Lipopolysaccharide Receptors; D017493:Leukocyte Common Antigens