Percoll density gradient centrifugation and consecutive flow cytometry do not identify leukocytes and leukocyte subtypes in ejaculate specimens.
This paper describes an attempt to establish a new combined method of leukocyte analysis in human ejaculate by Percoll density gradient centrifugation and consecutive flow cytometry. As a first step, leukocyte separation was performed by Percoll density gradient centrifugation with consecutive enrichment of leukocytes, especially granulocytes, in the 40%/60% and 60%/80% Percoll interfaces. Then these fractions were stained with specific monoclonal antibodies and analysed in a Facscan flow cytometer. Flow cytometric analysis did not demonstrate identifiable leukocyte populations, indicating a questionable cross-reaction with spermatozoal elements. Therefore, the combined technique of Percoll density gradient centrifugation and flow cytometric analysis were considered unsuitable for clinical leukocyte determination.
Diemer
T
T
Department of Urology, Georgia-Augusta University, Göttingen, Germany.
Weidner
W
W
Michelmann
H W
HW
Nierste
B
B
Ringert
R H
RH
D000911:Antibodies, Monoclonal; D002469:Cell Separation; D002499:Centrifugation, Density Gradient; D005434:Flow Cytometry; D006801:Humans; D007962:Leukocytes / Q000166:cytology*; D008297:Male; D012661:Semen / Q000166:cytology*
D000911:Antibodies, Monoclonal