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Andrology Lab Procedures

Introduction

Prior to entering an ART (Advanced Reproductive Technologies, e.g. IVF or GIFT) Cycle, a thorough evaluation of the quality and behavior of the spermatozoa must be completed. This requires performing several different assays that allow us to predict the ability of an individual's spermatozoa to fertilize an egg. Currently, however, there is no available assay to precisely determine the fertilization potential of a man's spermatozoa. Although these assays will help us develop a strategy to optimize fertilizing potential, they can not guarantee that fertilization will take place; compounding factors, such as egg quality may also have an effect on the rate of fertilization. The following assays must be performed prior to entering a stimulation cycle.

Semen analysis

A semen analysis is one of the simplest assays we perform. This assay provides us with baseline information regarding the quantity of spermatozoa in the ejaculate and the vigor with which they move. The numbers of normal spermatozoa in an ejaculate can vary tremendously from one ejaculate to the next and therefore, a minimum of two semen analyses must be performed. The first analysis will be scheduled during your orientation visit, and we will also schedule a follow-up analysis. Please read and follow the directions in the handout "Collection of Semen, Specimen Collection Instruction Sheet". The accepted normal values for men are as follows (World Health Organization, 1992):

  • 2.0mL or more
  • greater than 20 million spermatozoa/mL
  • greater than 40 million spermatozoa/mL per ejaculate
  • greater than 50% with slow/sluggish and rapid progressive motility or >25% rapid progressive motility
  • greater than 30% with normal forms
  • greater than 75% live spermatozoa
  • less than 1 million white cells/mL
  • less than 20% spermatozoa with antibodies bound

It takes approximately three months for a mature spermatozoon to form, and therefore any perturbation that occurs during this three month period may have an affect on the development of the spermatozoa. You should inform the fertility center physician of your activities during the previous three month period. Certain drugs may also affect sperm function (e.g. beta-blockers), so please let us know if you have been taking any medication during this time period.

It is sometimes customary to freeze semen for backup use during an ART cycle, in case the husband can not collect a fresh specimen. Since failure to collect is a very rare occurrence, you will be given the option of whether or not you wish to have a backup specimen frozen. More information is available here: 

Sperm antibody assay

An antisperm antibody assay must also be performed. This assay determines whether antibodies formed against spermatozoa are present either in the husband (a direct assay) or in the wife's serum (an indirect assay). We test the husbands living sperm directly, but test indirectly for antibodies produced by the wife. In the indirect assay, we incubate the husbands spermatozoa with the wife's serum, and then test for the presence of antibodies on the spermatozoa that were transferred from the serum.

We will collect a blood sample from the wife, usually during the orientation visit; this blood will be processed and frozen for later use. At this time we will also schedule the husband for a return visit to collect a second semen specimen. This second specimen will be used to perform a second semen analysis as well as the sperm antibody assay. We only perform antibody testing on Wednesdays, so it is essential to schedule a return appointment as soon as possible.

Additional assays

Dependant upon the results of the first two semen analyses, additional tests may have to be performed. These may include:

  1. Strict Criteria Assay. This is a cytological examination of spermatozoa after they have been stained and permanently mounted on a microscope slide. The examination consists of a critical assessment of spermatozoon morphology by a microscopic examination of 100 - 200 individual spermatozoa under high power. Recent studies have shown this assay to help identify individuals with low or negligible fertilizing potential. In cases where low fertilizing potential is identified, we can use certain strategies to help improve fertilization. Other specific ART techniques, such as spermatozoan microinjection must be considered in the worst cases.
  2. White Cell Identification. This assay specifically identifies white blood cells in the ejaculate. White blood cells should normally be present only in very small numbers in the ejaculate, and high concentrations are usually indicative of a reproductive tract infection. These cells are deleterious to spermatozoa and a urological consult may be required.
  3. Trial Swim-Up. In men with lower than average sperm counts or sperm vitality, this and additional assays will be required in order to determine whether sufficient numbers of motile sperm can be obtained to inseminate the eggs collected during an ART cycle. These assays include different sperm processing techniques that will allow us to develop the optimal conditions and methods to be used to maximize spermatozoon recovery.

For more information about treatment options

The Fertility Center
Geisinger Medical Center Women's Pavilion
Danville, Pennsylvania  17822-2920
(570) 271-7906

Geisinger Wyoming Valley Medical Center
Valley Medical Building
Wilkes-Barre, PA 18702
(570) 826-7914