Simultaneous flow cytometric detection of antibodies against platelets, granulocytes and lymphocytes.
We present a time-saving and objective flow cytometric immunofluorescence assay for the simultaneous detection of antibodies against platelets, granulocytes or lymphocytes using a reconstituted mixture of these cell populations. Platelets, granulocytes and lymphocytes could be distinguished on the basis of their forward (FSC) and sideways (SSC) light scattering properties plotted on scales of 4 log orders. After setting FSC/SSC gates around the platelets, granulocytes and lymphocytes, the reactivity of the sera with the cell populations was determined by histogram analyses of immunofluorescence for each gate. The flow cytometric assay of reconstituted cell mixtures showed a strong, positive correlation with a reference microscopic immunofluorescence assay of separate cell suspensions. The reproducible procedures for the isolation and staining of the cells and the electronic stability of the flow cytometer permitted the use of the same gate and marker settings throughout the experiments. Consequently, the entire analysis of data stored in list mode could be performed using a keystroke, so that time consuming and subjective manual analyses were avoided.
Blood Transfusion Service, Dr. Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
K
K
Sintnicolaas
W
W
de Vries
R
R
van der Linden
J W
JW
Gratama
R L
RL
Bolhuis
D000906:Antibodies / Q000032:analysis*; D015703:Antigens, CD / Q000032:analysis; D001792:Blood Platelets / Q000276:immunology*; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006021:Glycophorins / Q000032:analysis; D006098:Granulocytes / Q000276:immunology*; D006801:Humans; D016130:Immunophenotyping; D008214:Lymphocytes / Q000276:immunology*
D000906:Antibodies; D015703:Antigens, CD; D006021:Glycophorins