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Sperm Testing

For initial testing, a standard semen analysis may be done. If it is abnormal, or if history suggests probable sperm issues, the more comprehensive male factor screen and trial wash may be suggested.

Semen Analysis

The complete semen analysis procedure performed in the Andrology Lab consists of several procedures starting with the collection of specimen, analysis of the volume and ending with morphology. The semen analysis procedure described here follows closely the format outlined in the World Health Organization (WHO) laboratory manual for the examination of human semen and sperm-cervical mucus interaction. 

It is suggested that the patient undergo semen analysis testing on at least 2 separate occasions for accuracy. Although there is considerable variation in performing semen analyses from one laboratory to another, the results when used together with other sperm testing results will be most helpful to the doctor in diagnosing male factor problems.  


Part 1: Basic parameters - Volume, pH, Count, Motility

Part 2: Strict Criteria Morphology with Sperm Deformity Index

Part 3: Hyperactivation motility

Part 4: Chromatin Status, an indicator of both DNA fragmentation and chromatin structure abnormalities

Part 5: Predominant form of abnormal sperm shape

The strict criteria morphology test reports the number of sperm with a normal shape matching a specific profile of a fertile sperm. When more than 4 % of the sperm fit the shape profile, then, there is a high likelihood of a pregnancy with assisted reproductive technology.

Male Factor Screen 

The Male Factor Screen is a panel of related tests that help to determine if male infertility is a problem in certain patients. Although male infertility encompasses a wide range of infertility disorders, the Male Factor Screen focuses on subtle yet severe male factors such as denatured or broken deoxyribonuclei acid (DNA) in the sperm cell.

Sperm submitted for the Male Factor Screen will be tested for (1) hyperactive motility (2) strict criteria morphology (3) chromatin-relaxed acridine orange DNA integrity test and (4) the mini hamster test or sperm penetration assay. The first test will tell the physician if the patient’s sperm show hyperactivation, a special type of fertility dance movement. This movement is seen in sperm with the capacity to fertilize eggs. To optimize this test, the sperm cells are washed in special culture medium and stressed by exposure to high temperature before testing. Sperm specimens that show signs of hyperactivation are considered fertile.

The strict criteria morphology test reports the number of sperm with a normal shape matching a specific profile of a fertile sperm. When more than 4 % of the sperm fit the shape profile, then, there is a high likelihood of a pregnancy with assisted reproductive technology.

The head of the sperm is packed with DNA that when combined with the egg DNA completes the full set of chromosomes for a human being. When a sperm cell starts to die or undergo apoptosis, or, the sperm making process is incomplete, then, the DNA in the sperm head begins to break down by unwinding or fragmenting. This breakdown process can be detected using the DNA integrity test. In addition, the status of sperm DNA chromatin condensation is also checked using another test, the acidic aniline blue assay. Sperm specimens that are below normal may require intracytoplasmic injection (ICSI) and/or transfer of an extra embryo. Ask your physician about antioxidants, vitamins, folic acid and supplements to improve sperm DNA.

The mini hamster test measures the number of sperm penetrating and forming swollen heads inside specially prepared hamster eggs. The higher the number of sperm forming the swollen heads inside the eggs, the greater the fertilizing capacity of the sperm. Specimens that show many swollen sperm heads may not require the ICSI procedure. The results of the four tests combine to make the Male Factor Screen which will help your physician decide on the appropriate treatment for you.

Trial Wash

The sperm must be washed prior to intraunterine insemination. The ejaculatory fluid contains high concentrations of prostaglandins, a hormone secreted from the prostate gland. Prostaglandins are powerful stimulators of muscle contractility, and if introduced into the uterine cavity, would cause very severe cramping and pain.

The trial wash or sperm survival test is a reasoned approach to find the best method of wash for your sperm. Each individual's sperm will respond differently to the various sperm washing methods. During a trial wash, your sperm specimen is divided and washed using four different methods:

(1) the swim-up

(2) the two-layer colloid

(3) TYB or Test-Yolk Buffer

(4) a wash incorporating the motility stimulant pentoxifylline

The number of sperm cells recovered after each of the four washes is calculated and compared. Important parameters such as the percent motility, the percent progressive, and the percent sperm hyperactivation are analyzed by means of a computerized, automated sperm analyzer. Finally, sperm taken from each of the four sperm washes are stressed at 40 o C (a high temperature for sperm) and the percentages of sperm surviving the treatments are reported. The sperm survival test (SST) indices for the four washes provide information on the potential fertility of the sperm. An optional second semen collection may be requested from you to find out if a second specimen will help to increase the sperm count or concentration for treatment procedures such as artificial insemination or in vitro fertilization (IVF).

Interpretation of Results

For artificial insemination, the physician would like to have five million or more motile sperm cells after sperm washing, although pregnancies have resulted from lower sperm counts. For IVF without micromanipulation, we would like to recover greater than 500,000 motile sperm demonstrating about 5% hyperactivation and 75% or more of the sperm surviving the heat stress. Sperm of some patients do not respond favorably to stimulants such as pentoxifylline. The trial wash helps to reveal subtle sperm sensitivities before they are used on the actual day of insemination. The higher the percentage of sperm with hyperactive motility, the better the quality of the sperm specimen. Sperm showing 10% or higher percentages of hyperactivation are correlated to a higher fertilizing capacity.

Another important change that sperm must undergo before they can fertilize eggs is a process called capacitation. Capacitation generally peaks several hours after washing away seminal plasma components from the human sperm. Hence, in the trial wash, observation of the sperm cells several hours after washing helps to show which wash procedure maximizes the fertility potential of your sperm. It has been reported that fertile sperm can survive high temperature stress and it is expected that about 75% or more of the sperm will survive the stress. Sperm surviving the high temperature stress have been predictive of pregnancy after assisted reproductive techniques. Since semen specimens may vary in performance on different occasions, sometimes it is necessary to repeat the testing.