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Updated: Mar 7, 2026

Flow Cytometric Analysis of Biomarkers for Detecting Human Sperm Functional Defects
Published on: April 21, 2022
Validation of image cytometry for sperm concentration measurement: Comparison with manual counting of 4010 human
Dorte L Egeberg Palme1, Trine Holm Johannsen1, Jørgen Holm Petersen2
1Department of Growth and Reproduction, Rigshospitalet, University of Copenhagen, Denmark; International Center for Research and Research Training in Endocrine Disruption of Male Reproduction and Child Health (EDMaRC), Rigshospitalet, University of Copenhagen, Denmark.
Abstract:
Sperm concentration is an essential parameter in the diagnostic evaluation of men from infertile couples. It is usually determined by manual counting using a hemocytometer, and is therefore both laborious and subjective. We have earlier shown that a newly developed image cytometry (IC) method may be used to determine sperm concentration. Here we present a validation of the IC method by analysis of 4010 semen samples. There was high agreement between IC and manual counting at sperm concentrations above 3mill/ml and in samples with concentrations above 12mill/ml the two methods can be used interchangeable. However, we found substantial differences in samples below 3mill/ml. We also assessed the accuracy of the two methods by repeated measurements of 248 samples, which revealed that IC measurements seemed more accurate. Moreover, based on ten samples counted by several operators the IC method had a lower coefficient of variation than the manual method (5% vs 10%), indicating a better precision of the IC method. In conclusion, measurement of sperm concentration by IC can be used at concentrations above 3mill/ml and seems more accurate and precise than manual counting, making it an attractive option in the daily clinical practice.

