A sperm sample is best produced by masturbation. Lubricants cannot be used as they interfere with fertilization. To minimize the possibility of specimen contamination, washing of hands is crucial. Some males may find it problematical to produce a sample on request, and it is therefore important that this be discussed with the treating physician who suspects that this may occur.
For the most accurate analysis, the specimen should either be collected at, or reach the laboratory, within 1 hour of collection. If this is not possible, a specimen of up to 2 hours could be satisfactory. It should be collected into a sterile bottle and maintained at body temperature until it

reaches the laboratory. If required, it is also possible to freeze sperm on collection, and thaw it on the day of insemination. In view of the fact that the freeze/thaw process may cause damage to the sperm, it is not an option for marginal, or substandard quality sperm.
Use of drugs, alcohol, cigarettes or chewing tobacco should be kept to an absolute minimum during the 3 months before initiation of the treatment cycle. In some cases, the treatment may need to be postponed if a herpes lesion is present at the time of semen collection. Sperm quality may be adversely affected if the male had suffered from a fever of 101°F (38.3oC) or higher within 3 months prior to treatment. Sperm count and motility could appear normal, but fertilization may not occur.

Difficulties in Sperm Collection

When problems arise, discussions with counselors acquainted with all the options may help to resolve these.

Partner Assistance
Treatment of reproduction usually involves the couple; therefore, collection of semen should be regarded as a joint effort.

Interrupted Intercourse
This is not the method of preference, as the sperm could be contaminated with the vaginal flora, and it may also be possible to lose some of the ejaculate.

Condoms should be specific in type, since regular condoms are coated with spermicides and chemicals that are toxic to sperm. After intercourse the contents of the condom are emptied into a specimen bottle.

Retrograde Ejaculation
This occurs when the sphincter (valve) to the bladder fails to close and the semen, instead of being ejaculated from the penis, reverts to the bladder. This phenomenon could result from urological surgery and diabetes, but occasionally has no apparent cause. Since urine is basically acidic, it results in rapidly immobilizing as well as damaging the sperm. In order to solve these issues and produce viable sperm, urine, and especially the bladder environment needs to be changed to be basic (pH higher than 7.0). First, the male is frequently given a urinary alkalinizer in order to make the urine less acidic, and more basic. The male is then asked to empty his bladder, and then to produce sperm by masturbation. The collection of sperm is best done at the laboratory so that the processing of sperm washing is performed immediately after collection.

Spinal Injury Patients
Patients with spinal injury often have difficulties in obtaining penile erection and ejaculating. Semen specimens can be collected from these patients either by vibrator-induced ejaculation, or rectal electro-ejaculation. These patients may also have the complication of retrograde ejaculation, therefore the strategies outlined above, as well as filling and flushing the bladder with Tyrodes solution, are also employed.

Surgical Sperm Collection

Over the last few years, the field of reproduction has developed dramatically. One of the greatest achievements was the introduction of the intracytoplasmic sperm injection (ICSI) facilitating fertilization of the egg with one sperm that is injected into the oocyte. This option of fertilization opened a new field in medicine related to sperm retrieval or aspiration from the testis.
The purposes of surgical sperm collection are: a) to obtain the best quality sperm, and b) to have enough sperm for future procedures as well i.e. to cryopreserve the unused sperm; c) to minimize damage to the male reproductive system.

Testicular Fine Needle Aspiration (TFNA)

This technique was initially developed as a diagnostic procedure to investigate men with azoospermia. Percutaneous puncture and aspiration of the testis can be performed using a 21-23-gauge needle connected to a 20cc syringe, or with an automatic biopsy gun. Generally, sperm can be collected from males with obstructive azoospermia. The procedure may be performed with local anesthesia without any necessity for surgical involvement of the scrotum.


Male reproductive organs (After Dee McLean; In "Patient Pictures: Fertility" by Rod Irvine,
Published by Health Press, Oxford, 1966)

Normal testis

Percutaneous Epididymal Sperm Aspiration (PESA)


This technique can be performed without surgical scrotal exploration can be easily repeated, and does not require an operating microscope or expertise in microsurgery. The procedure can be carried out under local anesthesia. Usually the testis is stabilized between the surgeon's thumb and a 21-gauge butterfly needle attached to a 20-ml syringe, is inserted into the caput epididymis, and withdrawn gently until fluid can be seen entering the butterfly needle tubing. The procedure is repeated until an adequate number of sperm is retrieved.

spermcollection4 spermcollection5
Percutaneous Epididymal Sperm Aspiration  (PESA)

Percutaneous biopsy of the testis (PercBiopsy)
This technique is the most valuable for retrieving a large number of sperm in patients with obstructive azoospermia. A 14-gauge automatic biopsy gun is used under local anesthesia to remove a small segment of testicular parenchyma.

Microsurgical Epididymal Sperm Aspiration (MESA)
This is open surgery under the operating microscope. Individual tubules of the epididymis are isolated and micropuncture aspiration is undertaken. This approach has the advantage of reliable retrieval of large numbers of epididymal spermatozoa that can be readily frozen and thawed for subsequent attempts at fertility. Puncture sites are closed with interrupted nylon suture, or cauterized and the epididymal tunic closed.


Each sperm sample retrieved is immediately examined by an embryologist under a phase contrast microscope at 200 x power to evaluate the concentration and motility of the spermatozoa, as well as the degree of blood cell contamination.
Retrieval of sperm from the epididymis of males with obstructive azoospermia is possible in over 99% of patients.

Testicular Sperm Extraction (TESE)

A technique is available in which a biopsy is taken from the testes, the sperm collected and observed under the microscope. The technique requires multiple blind testis biopsies, with excision of large volumes (>700 mg) of testicular tissue with risks of permanent damage to the testis. A relatively avascular region of the testis needs to be located for an incision.





Testicular biopsies Testicular Sperm Aspiration (TESA)

The site is not a replacement for professional medical opinion, examination, diagnosis or treatment. Do not delay seeking or disregard medical advice based on information written by any author on this site. No health questions or information on is regulated or evaluated by the US Food and Drug Administration or any other administration, and therefore the information should not be used to diagnose, treat, cure or prevent any disease without the supervision of a medical doctor. Posts made to these forums express the views and opinions of the author only, and serve as an open forum to discuss clinical issues among experts in the field.