Next day determination of ejaculatory sperm motility after overnight shipment of semen to remote locations.
OBJECTIVE
To develop a method for delayed assessment of sperm motility, after shipment of semen to a remote laboratory. Sperm in semen were labeled with the MitoTracker(®) Red CM-H(2)XRos reagent, and fixed with 3.7 % formaldehyde by the laboratory technicians at the origin of the semen. This treatment reflected well sperm mitochondrial activity, and the MitoTracker(®) signal was related to sperm motility and velocity for 2-3 days following ejaculation.


METHODS
Sperm motility and velocity were evaluated manually and by computer assisted semen analysis (CASA), respectively. Fluorescence assessment of individual sperm was carried out with the computer assisted Metamorph v4.6.9 program. Emission levels of MitoTracker(®) spermatozoa were studied in room temperature and cooled semen, or in the respective room temperature swim-up sperm fractions following ejaculation, and on the second day (N = 103 samples, 89 men) and third day (N = 10 samples, 8 men).


RESULTS
Sperm with optical density (O.D.) ≥0.7 showed close correlations with ejaculatory sperm motility and velocity even after second day (r = 0.92, p < 0.001, N = 103 samples). Further, the multiple of sperm motility and velocity was also related to the proportion of high MitoTracker(®) reagent emission sperm (r = 0.83, p < 0.001, N = 103 samples). MitoTracker(®) dye fluorescence on the second day accurately reflected the ejaculatory sperm motility (r = 0.90, p < 0.001). Thus, a shipping delay would not adversely affect the results.


CONCLUSIONS
The delayed assessment of sperm motility in samples treated with MitoTracker(®) Red CM-H(2)XRos reagent and shipped to remote laboratory truly reflects the level of sperm motility at the time of the ejaculation.
Department of Histology and Embryology, Akdeniz University School of Medicine, Campus, 07070, Antalya, Turkey, leylasati@yahoo.com.
Leyla
L
Sati
David
D
Bennett
Michael
M
Janes
Gabor
G
Huszar
D015925:Cryopreservation; D004542:Ejaculation / Q000502:physiology; D006801:Humans; D008297:Male; D012661:Semen / Q000502:physiology*; D055101:Semen Analysis; D012662:Semen Preservation / Q000379:methods*; D013076:Sperm Count; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa