Arizona Andrology Laboratory and CryoBank: Complete Semen Analysis

Arizona Andrology Laboratory & Cryobank

Providing state-of-the-art male fertility testing, processing, cryopreservation, and storage

Gateway Wellness Center - 850 N. Kolb Road, Tucson, AZ 85710

Phone: 520-885-2689 - Fax: 520-731-0715

Complete Semen Analysis

The Complete Semen Analysis is the primary test performed to inform and guide the clinician in the evaluation of male fertility. It is important for the sample to be collected into a sterile container by masturbation after 2 to 5 days of abstinence and examined within one hour of collection in order to obtain the most accurate result. Time of collection and days of abstinence are noted on the specimen container label. The volume, color, odor, consistency, and pH are checked after the specimen is transferred to a test tube. A small amount of the semen is placed in a Microcell™ or Standard Count counting chamber to calculate the sperm density. For extremely low densities, the hemacytometer counting chamber may be utilized to quantify the count. Motility, forward progression, clumping, aggregation, rounds cells per high power field, debris, an estimation of morphology, and the presence of any bacteria may also be determined from the counting chamber or from a direct wet mount slide.

The Arizona Andrology Laboratory & Cryobank also has the capacity to film and archive live video images of the sperm specimens which can be an aid in the interpretation of the semen quality. Patients can also view the specimens on a monitor. In many cases a picture, or even better a video, is worth more than words or numbers can say.

The Complete Semen Analysis is usually one of the first and most common test ordered to check a man’s fertility. Unfortunately, not all labs examine or report out all semen parameters in the same way. The following is a guideline that our Laboratory uses in evaluating a semen sample that will hopefully remove some of the mystery:

The four critical parts of a semen analysis:

Parameter

Units

What it measures / How

Normal Range

Volume

How much “liquid” volume is in the sample.

The volume is read off the markings on a 15 ml conical tube.

1.5 to 5.0 ml

Density

M/ml

The number of sperm (in millions) in a volume of 1 ml of the sample. Sometimes called “the count”.

A counting chamber, like the Microcell, is used calculate the density from a small drop of the sample.

Semen washes can increase the density by concentrating the sperm into a smaller volume that may then be used for an IUI, or IntraUterine Insemination, a common and relatively inexpensive procedure that may be very helpful in increasing the chances for a couple achieving pregnancy.

50.0 to 120.0 M/ml

Motility

The percentage of sperm seen that have some movement or motion.

100-200 sperm are examined. If 35 out of 100 have motion, the motility is 35%, etc.

The percent motility can be increased in a sample being prepared for IUI by using an Isolation gradient wash that “filters out” many of the non-motile sperm, as well as other undesirable components of the semen sample.

50% to 80%

Forward Progression

FP (0 to 4.0)

An overall average of how fast and straight the motile sperm are swimming on a 0-4 grading scale.

The FP is a subjective estimate of the overall motion of the motile sperm from an examination on the slide.

2.5 to 3.5

The Forward Progression (FP) grades and what they mean:

Since the Forward Progression rating is probably the most subjective of the four critical parameters, here is a breakdown of what each rating means and how they are estimated. Note that a FP of 0 or 1.0 is not subjective.

0	The motility is 0%. None of the sperm are motile or moving. If the motility is 0%, the FP will automatically be 0. Not subjective.

1.0	Some of the sperm are motile, but none of them are moving ahead or showing any signs of forward motion. They are basically just “moving in place”. Not subjective.

1.5	A few of the motile sperm are making some forward motion or progression, but it is very slow. The rest of the sperm are either non-motile, or have a FP = 1.0.

2.0	Most of the motile sperm are moving forward in a slow or sluggish manner. A few might have a FP = 1.5 or a 2.5, but the majority are moving slowly.

2.5	The majority of the sperm have good, normal rapid forward progression. A few might have a FP = 2.0 or even 3.0, but most simply have normal progression.

3.0	Most of the sperm have a very rapid forward progression.

3.5	Most of the sperm have extremely rapid forward progression and appear to be hyperactive. A FP of 3.5 is usually only seen after a specimen has been washed.

4.0	All of the motile sperm have extremely rapid forward progression. Very rare, and usually only associated with sperm that have been washed.

In short, a Forward Progression grade of 0, 1.0, 1.5, or 2.0 is ABNORMAL, while a rating of 2.5 or better is considered normal. Semen processed with an Isolation Wash usually have sperm with better forward progression in the final IUI preparation since many of the slower or non-motile sperm are “filtered out”.

Other important semen analysis parameters:

Days of Abstinence (DAB)

Should be between 2 to 3 days – no more than 5 days.

Longer or shorter periods can affect all the critical numbers above, usually making them lower.

Hyperviscosity (HV)

Semen should liquefy within 1 hour after collection. The lab rates the viscosity or thickness of the sample on a scale of 0 to 4 (0,1,2,3,4), rather than a liquefaction time.

A rating of 0 is normal. The sample is not hyperviscous.

Hyperviscosity can affect the reporting of density, % motility, and even FP.

NOTE: This makes the Time Of Collection for a sample very important, since it is compared to the Time Examined to see how long the sample had to liquefy before it was graded.

Clumping

Indicates non-motile sperm than are clumped together. Also graded on a scale of 0 to 4 (0,1,2,3,4).

A rating of 0 or 1 is considered normal (some minor clumping is not unusual).

Clumping can affect the reporting of density and motility. Motile sperm caught in clumps are said to be aggregated. Isolation washes may remove clumps – Chymotrypsin washes may break up the motile sperm caught in clumps.

Round cells (RC)

Round cells are just that – cells other than sperm than are basically round in shape. They may be White Blood Cells (WBC’s), immature sperm or perhaps other cellular components. They are all grouped together as round cells since it really requires a special stain to differentiate the WBC’s from the rest, which is available as a separate WBC test. Round cells (RC) are reported out as a range seen per high power field (HPF), ie, 2-6 RC/HPF.

Anything less than 7 RC’s / HPF is considered normal.

A large number of round cells may indicate an infection (since many of the “round cells” may be WBC’s). Samples with large number of RC’s are also good candidates for isolation washes to help “clean up” the samples before an IUI.

Estimated Morphology

A general estimate of the percentage of apparently normal sperm forms seen. Somewhat subjective. If a large numbers of a particular type of abnormal forms is observed, these may be noted in the comments section of the report.

A rating of 50% or greater is considered normal.

To critically examine the morphology, sperm need to be stained and individually compared to a set of strict criteria. This is available as a separate test called the Strict Morphology (SM) assay. An isolation wash will generally improve the overall morphology of the sperm recovered, as well as the motility of the final sample ready for IUI.

Bacteria

Colonies can often be visualized even under 400X power, usually a tiny rods or cocci. Bacteria is noted in the comments section of a report if any are seen in the sample.

The presence of any amount of bacteria is considered abnormal.

Other parameters:

A pH of 7.2 to 8.0 is considered normal.

Debris

Includes anything observed in the semen that cannot be identified as sperm, round cells, or other common cellular components. Graded on a scale of 0-4. Some minor debris is common.

A grade of 0 or 1 is considered normal.

Isolation washes will remove most debris.

Color

Semen varies in color. It will usually be a pale yellow or somewhat opaque. Pink may indicate the presence of blood. A very “clear” sample may have low density.

Odor

A very strong or foul odor may indicate a long interval between time of collection and time delivered to the laboratory. Other factors can affect the odor.

Two Important Calculated Values:

Total Count Multiply Volume x Density. Ex: 2.0/45.0/65%/2.5 TC=2.0 x 45.0 = 90 Million. The total count is the total number of sperm in the entire specimen.

Total Motile Multiply Volume x Density x % Motility. Ex: TM = 2.0 x 45.0 x 0.65 = 58.5 Million. The Total Motile is perhaps the single most important value of the semen analysis. It is the total number of sperm in the entire sample that have some “swimming motion”. It is also the minimum number of sperm that are alive in the entire sample (other sperm may be alive, but not motile). The decision on whether the couple should consider an Intrauterine Insemination (IUI), the type of wash to use for the IUI, or a more advanced technique such as Itracytoplasmic Sperm Injection (ICSI) is often based on the total motile count of the semen specimen.

ArizonaAndrology.com.