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Semen Analysis usually is not given a lot of importance. It is very much dreaded by people who do it also and normally it is done in a very lack sidal manner. We feel that if we get an idea about what a sperm sample looks like, it will more than serve our purpose. But we need to give this analysis more respect than this step-child treatment because it is the basic diagnostic test of a man's fertility. There are several aspects to a basic semen analysis and we are going to cover them all in this blog.

The human semen is composed of only 5% spermatozoa. The rest are secretions from all the accessory reproductive glands. The bulbo-urethral gland contributes 10% of the semen, the prostate contributes 20-30% and the seminal vesicle, the major portion 46-80% which is alkaline in nature cancels out the acidity of the prostate because sperm cannot survive in an acidic environment. So, semen analysis is a pivotal evaluation in the diagnosis of the male infertility. A decrease in sperm concentration, motility or morphology is associated with reduction in fertility.

Semen analysis is also called semen gram. It evaluates the characteristics of the semen and sperm. Semen analysis has three parts to it. 

·       First is the pre-examination procedure,

·       second the actual examination

·       the post-examination procedure.

Pre-examination procedure is the patient instructions. You have to instruct the patient to have an abstinence of not more than three days. You have to tell him to come mentally prepared for the collection and maintain as much sterile environment as possible. Then sample correction, sample receipt from the patient to the lab and the initial handling. Labelling of sample is very very important because without labelling you will not know whose sample you are checking.

The examination procedure happens between the first 30 to 60 minutes. Some tests can happen within three hours and some can be performed later in the same day. But never a sample is kept overnight.

Post-examination procedures involve calculations, authorisation of results if required and the release of result to the patient.

Semen Collection

Abstinence should not be more than 3-5 days. The sample is collected into a clean wide sterile container which is provided to the patient by the lab. 70 percent of the sperm is in the first part of the ejaculate. The patient must be instructed strictly that he is not supposed to lose any part of the sample otherwise the analysis will not be correct. And as I mentioned labelling of sample is of paramount importance. Now some patients they cannot collect the sample in a clinical setup.

They prefer to collect at home or in an environment where they are more comfortable like a hotel or some place. Usually, it is advised to discourage this practise mostly because of handling and chain of custody. If a patient comes to you with a similar sample saying that it is his but in fact it belongs to somebody else then there is no way of proving that the patient is not telling the truth.

Home collection should be discouraged but if there is absolutely no way in which the patient is being able to collect sample in the clinical setting then within 30 to 60 minutes of collection the sample should come to the clinic. It should not be more than that.

Steps of semen analysis?

·       You collect the sample then place the sample in an incubator at 37 degrees for about 30 minutes.

·       After that you perform the macroscopic examination and the microscopic examination. So macroscopic examination within 30 to 60 minutes involves a liquefaction and appearance, the volume of the semen, the pH of the semen and wet preparation making. Wet preparation is basically put a drop on the slide and put a cover slip on top of that for microscopic examination.

·       And a microscopic examination constitutes of sperm concentration, sperm motility, vitality, morphology, mixed agglutination test that is for sperm antibodies and assessing peroxidase positive cells if you see a lot of WBCs present in the sample.

·       Now semen analysis, other tests that can be performed within three hours depend on the sperm concentration and within three hours the semen sample can also be sent for culture if you are suspecting any sort of contamination or presence of more leukocytes in the semen sample.

Later on the day you can assess the smears if you have already prepared the smears for the morphology of the sperm then you can check that later in the day because those sperms are fixed they are not going anywhere. Accessory gland markers like seminal vesicle and prostate glands that test can be performed later on and indirect immunobead test to check for anti-sperm antibodies. These tests can be performed later in the day not necessarily within 30 to 60 minutes.

Safety while handling biological samples is of paramount importance especially to the worker so semen sample may be infectious and it absolutely should be handled as a biohazard. In such cases, ensure you are being trained under the IVF Doctors in Bangalore. So, normally it is preferred that you tell the patient to get the triple marker test done before you are taking any sort of biological sample from him but the patient might still be in a window period and might not reflect in the test so any and all biological material that you use should be handled with absolute care and treated as hazardous. Lab staff must be immunised for hepatitis B and PPE if possible, should be used. Surface disinfection should be taken very seriously and all the disposables should be immediately discarded.

Liquefaction is a breakdown of the gel portion of the seminal plasma which is facilitated by the enzyme secretion from the prosthetic fluid. Abnormal sample takes more than 60 minutes after ejaculation. It might never liquefy. You can chemically treat the viscous samples to liquefy them by Dalbecker's phosphate, buffered saline, bromelain and chymotrypsin.

How do you check for cement viscosity?

You gently aspirate it into a wide bore plastic disposable pipette. Allow the cement to drop by gravity drop by drop and observe the length of any thread which is formed in between. So a normal liquefied ejaculate falls as small discrete drops, you know drop by drop it will fall down, but if viscosity is abnormal the drop will form a thread which is more than two centimetre long and it is very easy to identify if a sample is viscous or not.

The appearance of the sample, it is very important and gives you a lot of information. The normal cement sample is homogeneous and it is grey or pelicant in appearance. If there is a longer abstinence, then the sample might be slightly yellowish. If RBCs are present, if there is any injury, then red or brown colouration might be there. Yellow colour is an indication of jaundice or taking of certain drugs or vitamins and viscous which is totally clear and colourless, then it may be a pre-ejaculated only from the caupers gland.

So seminal vesicular secretion is mostly alkaline and prostate secretion is acidic. Both of these balances out to form a pH of about 7.2 to 7.4 of the seminal plasma. If you keep the sperm at room atmosphere for a longer time, then there will be a continuous loss of CO2 which will cause a gradual increase in pH. It will make the sperm more acidic. You can check the pH by using pH strips of 6 to 10 range. You have to perform analysis between 30 and 60 minutes of collection as soon as possible.

To find out how many percentages of sperm is abnormal, the percentage of morphology is a very time-taking procedure. You must manually count at least 100 sperm and out of that decide how many are normal and how many are abnormal and like this you must check multiple fields to get an average. So, this is a time-consuming procedure but it is worth it if you want to determine the morphology of the sperm because at the end of the day the sperm morphology can determine whether the patient will go for IVF or whether the patient is a candidate for ICSI

Semen Analysis is not as simple as it sounds in this blog, the knowledge goes way beyond this. If you are interested to learn in-depth about this interesting topic, then check your embryology course eligibility and join the next batch of Fellowship in Embryology at Medline Academics.

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Student enrolled for this program also gets a 3 week long clinical posting in Dr. Kamini Rao Hospitals for the practical training which covers every aspect of the male reproductive functions and test. This centre containing the best IVF doctors in Bangalore, is definitely recommended for training and treatment.

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