Flow cytometry and microscopic acridine orange test: relationship with standard semen analysis.
Improved prediction of male fertility requires advances in semen analysis. This study examined the reproducibility and independence of the flow cytometry acridine orange test (FCM-AOT) of sperm chromatin integrity as an assessment of semen quality. The study found that FCM-AOT results are not significantly affected by up to 6 h delay in semen preparation (n = 9) or contamination of semen with moderate concentrations of bacteria (<10(8)/ml E. coli or Staph. epidermidis, n = 14). The variation of replicate measurements within samples was low (%Abnormal alpha(t): SD = 1.4, 95%CI = 4.6, n = 25) and different samples from the same men were mostly within the range of measurement error (n = 35). FCM-AOT variables, in particular %Abnormal alpha(t), displayed significant correlations with motility (r = -0.557), vitality (r = -0.469) and morphology (r = -0.464, n = 201), which are similar in magnitude to those existing between the standard semen variables. Surprisingly, no correlation was found between %Abnormal alpha(t) and the microscopic acridine orange test (M-AOT) (n = 185), suggesting the FCM results are sensitive to a different aspect of sperm quality. In summary, this study confirms that although not totally independent of standard semen analysis or the M-AOT, it is found to be a robust, sensitive and reproducible measure of semen quality, representative of the individual.
Apedaile
Anwyn E
AE
Clinical Sciences Centre, Medical Research Council, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK.
Garrett
Claire
C
Liu
De Yi
DY
Clarke
Gary N
GN
Johnston
Stephanie A
SA
Baker
H W Gordon
HW
D000165:Acridine Orange; D002470:Cell Survival; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D006801:Humans; D007248:Infertility, Male / Q000175:diagnosis* / Q000473:pathology; D008297:Male; D015203:Reproducibility of Results; D012661:Semen; D012680:Sensitivity and Specificity; D013076:Sperm Count; D013081:Sperm Motility; D013094:Spermatozoa / Q000473:pathology*
D005456:Fluorescent Dyes; D000165:Acridine Orange