Morphometric dimensions of the human sperm head depend on the staining method used

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dc.contributor.authorMaree L.
dc.contributor.authorDu Plessis S.S.
dc.contributor.authorMenkveld R.
dc.contributor.authorVan Der Horst G.
dc.date.accessioned2011-05-15T16:16:33Z
dc.date.available2011-05-15T16:16:33Z
dc.date.issued2010
dc.description.abstractBackground Assessment of sperm morphology (including morphometry) is extensively used to determine one of the qualities of a semen sample and depends on the differential staining of spermatozoa. A staining technique should cause as little change to sperm dimensions and form as possible in order to reliably evaluate the morphometric features of the sperm. Various staining techniques have been employed, but only a few have been recommended by the World Health Organization and are amenable to automated sperm morphometry analysis. Our study was aimed at comparing the effect of three staining techniques [Papanicolaou (PAP), Rapidiff® (RD) and SpermBlue® (SB)] on human sperm head dimensions and to compare these with the head dimensions in fresh semen. Methods Smears made from human semen samples (n = 24) were stained according to the three staining techniques and sperm head morphometry was assessed with the Sperm Class Analyzer. Head dimensions of fresh spermatozoa were measured with a digital calliper on a computer screen. The minimum number of spermatozoa to be analyzed to represent the sperm population and the degree of inter-laboratory variation were determined. Electron micrographs from the same semen samples were used to determine the actual acrosome coverage of the spermatozoa in the semen (n = 7) in order to verify the Results of the automatic analyses. Result SThe osmolality of human semen differs from that of the RD and PAP fixatives and stains, but is more similar to the SB fixative and stain. At least 100 spermatozoa should be analyzed to include a representative sample of the sperm population. RD caused sperm heads to swell, PAP caused them to shrink and SB had no significant effect on sperm head dimensions when compared with spermatozoa in fresh semen. Very little inter-laboratory variations were found. The percentage acrosome coverage was significantly different between the three staining techniques, as well as between the RD and PAP stains and the manual measurements obtained using the electron micrographs. Conclusions Different staining techniques change the morphometric dimensions of the human sperm head, probably due to the fact that either the fixatives or stains are not iso-osmotic in relation to human semen. Since these changes in sperm head dimensions are not uniform, care should be taken when selecting a staining technique. Ideally, stained spermatozoa should have dimensions as close to spermatozoa in fresh semen as possible, as was found with the SB staining method, Resulting in accurate evaluations of sperm head morphometry. © The Author 2010.
dc.description.versionArticle
dc.identifier.citationHuman Reproduction
dc.identifier.citation25
dc.identifier.citation6
dc.identifier.issn02681161
dc.identifier.other10.1093/humrep/deq075
dc.identifier.urihttp://hdl.handle.net/10019.1/13832
dc.subjectfixative
dc.subjectacrosome
dc.subjectanalyzer
dc.subjectarticle
dc.subjectcomputer
dc.subjectcontrolled study
dc.subjectelectron microscopy
dc.subjecthuman
dc.subjecthuman cell
dc.subjectintermethod comparison
dc.subjectmale
dc.subjectmorphometrics
dc.subjectosmolality
dc.subjectPapanicolaou test
dc.subjectsemen analysis
dc.subjectspermatozoon
dc.subjectspermatozoon count
dc.subjectstain
dc.subjectstaining
dc.subjectAnalysis of Variance
dc.subjectCell Shape
dc.subjectHumans
dc.subjectMale
dc.subjectMicroscopy, Electron, Transmission
dc.subjectSpecimen Handling
dc.subjectSperm Count
dc.subjectSperm Head
dc.subjectSpermatozoa
dc.subjectStaining and Labeling
dc.subjectStatistics, Nonparametric
dc.titleMorphometric dimensions of the human sperm head depend on the staining method used
dc.typeArticle
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