Sperm Morphology by High Magnification in Fertility Men
Sperm Morphology by High Magnification in Fertility Men
A new concept for observing the fine morphology of spermatozoa at high magnification (x6000)
 with an inverted microscope, a numeric camera using differential interference contrast has
 been developed (1). This technique called Motile Sperm Organellar Morphology Examination
 allows to see some abnormalities, mainly vacuoles on the head of spermatozoa. These vacuoles
 appear to be related to sperm DNA damage and to affect embryo developmental potential (2, 3,
 4). The application of Motile Sperm Organellar Morphology Examination may represent an
 improvement in the evaluation of semen quality, with some potential clinical repercussions at
 the diagnostic/prognostic level. First of all, the investigators need data on fertile men in
 order to define " normality " of sperm morphology at high magnification. The aim of this
 study is therefore to better characterize these vacuoles (number, surface, position) in a
 population of men fertile in order to establish normality criteria.
The population studied consisted in 50 men aged 18 to 45 years with proven spontaneous
 fertility. All subjects gave their informed consent to participate in the study. After
 questioning on full medical and andrological history, semen samples were collected by
 masturbation after 2 to 5 days of sexual abstinence and were processed for analysis after
 liquefaction for 20 min at 37°C. We carried out a sperm count, motility, vitality and
 conventional morphology analysis as well as a detailed morphometric analysis of the vacuoles
 at high magnification using an image analysis software. For the analysis at high
 magnification, fifty microliters of fresh sperm was washed in 2.5 ml of washing solution by
 centrifugation for 5 minutes at 400g. The pellet was resuspended in 100 µl of washing
 solution and the spermatozoa were fixed by addition of 100 µl Phosphate Buffer
 Saline-formaldehyde 3.7%. Two microliters of this suspension was placed in a glass-bottomed
 dish and examined by Nomarski interference contrast microscopy with a camera mounted on a
 microscope with an immersion objective lens x100. For each sample, sperm head vacuoles were
 analyzed on 100 spermatozoa that were randomly photographed and separately analyzed using
 digital imaging system software. Measurements using the software were carried out by a single
 operator. The Interactive Measurement module allows measurement of sperm head areas and
 vacuole areas by manually depicting their outline. The area and position of each vacuole were
 recorded.
INCLUSION CRITERIA:

 -  Age : between 18 and 45 years

 -  Sex : male

 -  Men with proven spontaneous fertility (the pregnancy started less than two years
 before inclusion)

 -  2 to 5 days of sexual abstinence

 -  All subjects gave their informed consent to participate in the study

 -  The subjects must be affiliated to a social security scheme.

 EXCLUSION CRITERIA:

 -  Personal history of infertility

 -  Personal history of chemotherapy

 -  History of recurrent miscarriage in the couple

 -  Andrological history since the last pregnancy: urogenital infection, testicular
 trauma, testicular torsion, varicocele, abdominopelvic surgery

 -  Fever within 3 months before inclusion.