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

Cytometry A. 2006 Mar;69(3):127-30. doi: 10.1002/cyto.a.20216.

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.

MeSH terms

  • Antibodies / chemistry
  • Antibodies / immunology
  • Antigens, CD19 / analysis
  • CD3 Complex / analysis
  • CD4 Antigens / analysis
  • CD8 Antigens / analysis
  • Fluorescent Dyes / chemistry
  • Humans
  • Image Processing, Computer-Assisted
  • Immunophenotyping / methods*
  • Laser Scanning Cytometry / methods*
  • Leukocyte Common Antigens / analysis
  • Leukocytes / chemistry
  • Leukocytes / cytology
  • Leukocytes / immunology
  • Lipopolysaccharide Receptors / analysis
  • Lymphocyte Subsets / chemistry
  • Lymphocyte Subsets / cytology
  • Staining and Labeling / methods*

Substances

  • Antibodies
  • Antigens, CD19
  • CD3 Complex
  • CD4 Antigens
  • CD8 Antigens
  • Fluorescent Dyes
  • Lipopolysaccharide Receptors
  • Leukocyte Common Antigens