With a little practice you will become quite proficient at doing your own semen analysis.
To do a sperm count and evaluate motility and morphology you will need:
For truly accurate sperm counts you will also need Standard Depth Slides. You can see our microscopes here. We provide links to semen analysis supplies so you can easily order the equipment you need.
Do Your Own Sperm Count
Become Familiar with the Microscope
Set up the microscope somewhere where you can be comfortable. Review the microscope instructions and become familiar with the microscope's coarse and fine focus adjustments.
Start with the Stained Sperm Slide
Figures 1 and 2 show stained sperm slides. This lets you see what sperm look like under your microscope. (Clicking on an image at right launches a popup window with a larger view.)
Starting with the stained sperm slide helps you get familiar with the microscope and lets you see what sperm look like under your microscope.
Focus the Microscope
The large knobs on the microscope are the coarse focus adjustment. The small knobs control the fine focus. (The Model 107 LED does not have a fine focus.) Focus with the coarse focus knobs using the 10X objective. Then, zero in with the fine focus. Switch to the 40X objective. Refocus to get a clear image. Note about how big the sperm are with the 10X and 40X objectives.
If You Have Trouble Focusing
If you cannot see anything at any power it is probably because the fine focus adjustment is preventing you from moving the stage up far enough, or the barrel down if using a Model 109 or 107. To correct this try moving the fine focus adjustment to the far end of its travel in both directions. This can take many revolutions of the fine focus knob, keep turning. At each end of the fine focus travel, use the coarse adjustment to focus. After you get the microscope focused with the 10X objective, rotate the 40X objective into place. The 40X objective has a blue ring. Sharpen the image with the fine focus. Now adjust the Iris or rotate the Disc Diaphragm to see the effects of different amounts of light.
Three to Four Days of Abstinence
To get an accurate sperm count you should have three to four days of abstinence before collecting the sample. Note that longer is not better. The sperm tend to die off if you wait longer than four days. Some have found that shorter periods of abstinence give them higher counts and better motility.
It is fine, and helpful, to check things out without worrying about three to four days of abstinence. One of the nice things about having your own microscope is you do not have to do everything perfectly each time you want to have a look. It is OK to collect a sample after intercourse. It is helpful to check out motility and get a sense of the number of sperm present without the trouble of masturbating or having three or four days of abstinence.
Do a Sperm Count
Collect the sample by masturbation or using a semen collection condom after three or four days of abstinence. Let the sample sit at body temperature for about 1/2 hour before proceeding. You can put the sample inside your shirt to keep it at body temperature. This allows the sample to liquefy.
Use a pipette to apply a small drop of the semen sample to a microscope slide. Gently place a cover slip over the drop. Try to avoid getting air bubbles. Avoid finger prints by holding the slide and the cover slip by their edges. Be sure to use only one cover slip. The drop should spread out to just fill the cover slip. If it oozes out from under the cover slip use less. If it under fills the cover slip use more. It’s OK to waste a few slides to get this right. This gives you a sample depth of approximately 20 microns.
You can buy Standard Depth Slides with an attached cover slip that creates a fixed depth of 20 microns. This is the way to get very accurate, reproducible counts. These are what commercial laboratories use to do sperm counting.
Put the slide under the microscope. Focus as described above. Count the number of sperm that you see in one field of view when using the 40X objective. (If there are a lot you can count them in a quarter of a field of view and multiply by four.) Move the slide around and count 10 to 20 fields of view. Take the average of the counts. If you do not see any sperm go back to the human sperm slide to be sure you are focusing correctly.
Convert to Million Sperm per mL
Take the average number of sperm per field of view and multiply by 0.5 if using a Model 109 or Model 107 LED. Multiply by 0.3 if using a Model 131-CLED, 155, or DC-128 microscope. This gives your sperm count as the number of sperm in million per mL. A sperm count of more than 20 million per mL is good. See our What is Normal page to see how your results compare.
If the sperm are moving so much that you cannot count them you can put the sample container in hot water to kill the sperm. If you can find it, adding a drop or two of formaldehyde also works to immobilize the sperm. For the field of view on our microscopes, if there are so many sperm and they are moving so fast that it is hard to count them you have no worries.
Total Sperm Count
Now take your graduated cylinder and measure the total volume of semen. Multiply the volume in mL by the number of sperm in million per mL to get the total number of sperm.
Motility describes how much the sperm are moving. For sperm to do their job they need to be good swimmers. You can have good sperm counts but if there is no motility pregnancy is unlikely.
Sperm motility is segregated into four classes. The classes are:
- Rapidly progressive– swimming rapidly, generally maintaining a consistent direction
- Slowly progressive– moving slowly with some forward progression
- Non-progressive– thrashing about but not going anywhere or going in circles
- Immotile– not moving
You should evaluate the motility of a total of 200 sperm. You should do it twice for each field of view. In total you will have evaluated 400 sperm. First count the rapidly and slowly progressive sperm. Then count the non-progressive and immotile sperm. Calculate the average percentage of sperm for each class. The total percentage should be 100%. Enter this in the spreadsheet if you are using it. As a general rule you want more than 50% progressive sperm.
Evaluating morphology is easier if you use Pre-Stained Morphology Slides. These slides are used for the staining of spermatozoaand and are typically used to help the technician see the sperm for a morphology assessment. Each slide contains a dried layer of stain that makes it easier to clearly see the sperm. You can evaluate morphology without using pre-stained slides it is just not as easy.
To use Pre-Stained Slides, follow these steps:
1. Apply a small drop (approximately 2-4 microliters) of semen on the slide (Figure 1). If you are careful you can get two analysis out of each slide by using the left and right sides of the slide.
2. Place a cover slip on top of the slide, exerting slight pressure on the cover glass (Figure 2).
3. Allow slide to stand at room temperature for a few minutes before microscopic observation is made.
There are several classes of sperm defects including head, neck, mid-piece, and tail defects. Head defects include large, small, tapered, pear-shaped, round, amorphous, and double heads. “Pin head” or “micro head” sperm should not be counted. Neck defects include bent tails where the mid-piece and tail form an angle of greater than 90 degrees to the long axis of the sperm head. Other neck defects include asymmetrical tail insertion and thick or thin mid-piece. Tail defects include short, double, hairpin, broken, bent (more than a 90 degree angle), irregular width, or coiled tails.
Add it all up– how does your semen stack up?
The Merck Manual of Diagnosis and Therapy gives the following guidance on what is generally considered a measure of acceptable fertility after the ejaculate has liquefied for 20 to 30 minutes. The following characteristics represent adequate fertility:
- Ejaculate volume - 2.0 mL or more
- Viscosity - Liquefies within 1 hour
- Gross and microscopic appearance - Semen should be opaque or cream colored
- Sperm count > 20 million per mL
- Sperm motility at 1 and 3 hours > 50%
- Sperm morphology > 60% are normal
This guidance should not be considered absolute. These values do not represent the minimum values needed for conception. Many couples get pregnant with far lower sperm counts and poor motility. Some researchers have suggested that the fertility “cutoff” levels should be lower than the values listed above. If your levels are low take your vitamins, ejaculate frequently, and avoid hot tubs. See discussion on Improving Sperm Health.
You can see a table of results done with a microscope and by professional labs. We also offer a free spreadsheet you can download to do the calculations and to keep track of your results. The table at the bottom of the spreadsheet shows actual results. The table includes results for semen analysis done at home and semen analysis done by a clinic.
You can download a pdf of the 5th edition of the WHO laboratory manual for the Examination and processing of human semen.