Enumeration ofsemen leucocytes by fluorescence insitu hybridisation technique

Aim-To determinewhetherthefluorescentin situhybridisation technique (FISH)usingatotal humanDNA genomic probecanbeusedtoenumeratesemen leucocytes. Methods-Semen samplesfromfivedonorsweresubjected toamildKClsolution. Thesesampleswerethenbiotin labelled underFISH conditions usinga total humanDNA genomicprobeandtheleucocyte countsweredetermined. Tocheck theaccuracyofthetechnique a monoclonalantibodyagainstthe common leucocyte antigen CD45 [KC56(T-200)] servedasa control. An isotypic control for[KC56(T-200)], theinumunoglobulin [MsIgGl], served asasecondary control. Results-Semen leucocytes stained bythe FISHtechnique wereeasily detected becauseoftheir distinct bright yellow colour, whilethespermcells werered.Theleucocytecountrangedfrom0-5to49x106 permlofsemen.KC56(T-200) anditsisotypic control MsIgG1, whichserved ascontrolfortheFISHtechnique, accurately identified 94%and97%ofthesemenleucocytes ofacontrol donor, respectively. Conclusions-The FISHtechnique using atotal humanDNA probecanaccurately andeffectively enumeratetheoverall leucocytepopulation insemen. (JClin Pathol: MolPathol 1995;48:M319-M321)