Sperm Morphology by High Magnification in Fertility Men
Condition: Men; Fertility
Study type: Interventional

Summary: 
      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.
    

Detailed description: 
      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.
    

Intervention name(s): Assessment of sperm morphology (contrast microscopy).
Intervention type(s): Other
Intervention description(s): Sperm head vacuoles were analyzed on 100 spermatozoa that were randomly photographed and separately analyzed using digital imaging system software. 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. Relative vacuole area is the ratio of the area of all the vacuoles of a spermatozoon to the area of its head.
Intervention model: Single Group Assignment
Masking: None (Open Label)
Primary purpose: Diagnostic
Keywords: sperm morphology; vacuoles; fertile; Motile Sperm Organellar Morphology Examination; high magnification; reference values