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Diagnostic Semen Analysis

The diagnostic analysis of human semen and/or sperm can reveal answers to the cause of a couple’s infertility. If subnormal values are detected in the patient’s semen, then a diagnosis of male factor infertility may be listed as a contributing factor to the case. This diagnostic information pertaining to the quality of the semen samples is crucial to the Physician’s determination of the infertility treatment path. Routinely, new infertility patients will be requested to undergo a diagnostic semen analysis. Semen analysis results are determined according to the guidelines established by the World Health Organization (WHO).

Semen Analysis with Strict Morphology and Vitality Staining

These two advanced parameters are not included in diagnostic semen analysis all the time. Strict morphology of spermatozoa explains the detailed morphological abnormalities of sperm. This helps to find the causes of failed fertilization when semen having normal values is used for insemination.

Staining of sperm with vital stains helps to know the ratio of live and dead spermatozoa especially in case of low motility. Not all the immotile sperm are dead, staining of sperm differentiate between live and dead sperm.

For a review of how important sperm morphology is, please read the following fact sheet by ASRM Sperm morphology.pdf. Some men may have a varicocele that can impair fertility. Read more about varicoceles at: Varicocele.pdf

Semen Analysis after Sperm Wash

Semen analysis is done before and after washing of sperm with culture media. This will help to find out how sperm behaves during washing process. The procedure is indicated for future IUI patients.

Sperm Longevity Testing

Sperm after washing and removal of seminal plasma are cultured for extended period of time. The motility of spermatozoa is checked after 24 hours. A normal result is encountered when > 70% of the sperms are alive and moving at 24 hours.

Hypo-Osmotic Swelling (HOS) Test

This test is based on the semi-permeability of the intact cell membrane, which causes spermatozoa to swell under hypo-osmotic conditions, when an influx of water results in an expansion of cell volume. The HOS may be used as an optional, additional vitality test. This gives additional information on the integrity and compliance of the cell membrane of the sperm tail.

Retrograde Ejaculate Testing

Retrograde ejaculation involves reflux of spermatozoa in the backward direction into the bladder instead of the normal forward movement along the urethra to the exterior. This is often caused by an incompetent bladder neck. Some of common causes include: diabetes, iatrogenic surgical damage to the bladder neck innervation, and as a side effect of hypertensive therapy using alpha-adrenergic blockers. Clinically, men with retrograde ejaculation will present with either aspermia or no apparent ejaculate. In these cases diagnostic confirmation involves examination of the post-ejaculatory urine samples for the presence of spermatozoa.

Preparation of Fresh and/or Frozen-Thawed Sperm
for Intra-Uterine Insemination (IUI)

The rationale for the use of IUI is to reduce the effect of factors that may impede the progress of spermatozoa such as vaginal acidity and cervical mucus hostility, and to benefit from the deposition of a bolus of concentrated highly motile, morphologically normal sperm as close as possible to the oocytes. The use of washed prepared sperm for IUI has resulted in a significant reduction in the side effects associated with the use of undiluted semen for IUI, such as painful uterine cramps, and infection. For therapeutic IUI, a variety of sperm preparation methods are available. The most important consideration is to obtain a debris-free sperm suspension of adequate concentration and motility. The normal and motile sperm for IUI are separated by the Discontinuous Density Gradient and followed by repeated washing of the final suspension. Finally, the sperm pellet is mixed with a minute quantity of culture media, and then inseminated into the uterus using a special sterile catheter.

Cryopreservation of Semen

Cryopreservation of semen may be advantageous to couples in instances where the male partner is to undergo irradiation or chemotherapy, when sperm are retrieved surgically, or by electro-ejaculation, when the male partner is unavailable at the time of follicular aspiration due to previous commitment or situation, when the male partner is planning to undergo vasectomy, and/or is unable to produce on the day of follicular aspiration during IVF. The semen analysis is performed in the fresh ejaculate, after diluting with freezing media, and before and after the freezing process. The semen is then loaded in cryo-vials, and frozen in liquid nitrogen vapors. The frozen semen vials are then transferred very carefully into the liquid nitrogen tank (temperature -196°C) until used for IUI or IVF.

Dr. Fady Sharara

Dr. Fady Sharara Discusses Preimplantation Genetic Screening