Sperm collection explained

Professor Suks Minhas
Consultant Urologist and Andrologist , Imperial College London

Male Factor

From this video you will find out:
  • Is the fertility rate decreasing?
  • What are the main causes of male infertility?
  • What is a common reason for obstruction in males?
  • What are reconstructive options for obstruction?
  • What techniques are used for surgical sperm retrieval? When is it indicated?
  • What is the difference between TESE and micro TESE?
  • What is the role of optimizing testosterone before Microdissection Testicular Sperm Extraction (micro TESE)?

What types of sperm retrieval procedures are there?

In this session, Professor Suks Minhas, Consultant Urologist and Andrologist at Imperial College London, explained what the process of sperm collection looks like, how semen is analysed, what surgical sperm retrieval techniques are used and when are they indicated.

Sperm collection explained

Male infertility is important since 50% of infertility cases are due to male issues. It is defined as a failure to conceive after regular unprotected intercourse for at least 12 months. In addition to this, considering that infertility affects 15% percentage of couples, it is said that the male factor is responsible only for 20% of infertile couples and also contributes to 30% to 40% of cases. In this case, azoospermia, which is the non-existence of sperm in an ejaculation, is commonly related to these issues.

Causes of male infertility

On the one hand, It is acknowledged that a percentage of 30% of infertile patients are termed as unexplained infertility with no apparent cause. On the other hand, another series of causes can explain male infertility such as hormonal problems, sexual factors, and erection problems as well as considering infections.

  • anatomy of the male reproductive system

Sperm is produced in the testicles and then the production of sperm occurs in the epididymis, undergoing some changes that affect the ability of it to penetrate the egg to be later transported by the vas deferens, a structure known for conveying sperm directly into the body towards the seminal vesicles for ejaculation.

  • anatomy of the testicle and spermatic cord

In the male anatomy, the dilatation of varicose veins can cause heat damage, causing a lowering of sperm counts as well as affecting DNA cells.

Semen analysis

  • Standard collection
    • Abstinence (Between two or five days)
    • Collection method (Spermicidal)
    • Transport (Transportation to the laboratory at least in 1 hour)
  • Quality control
  • At least 2 samples

Azoospermia obstruction

In patients with azoospermia, the reason for this condition can be explained due to a blockage and also non-obstructing reasons as well. Most blockages are believed to come from the epididymis due to infections and trauma, but significant cases equally include the absence of sperm tubes or van deference. Low ejaculative volume is also found due to blockages near the prostate.

Vasal aplasia

Vasal aplasia also seems to be very important, as it causes the absence of the sperm tubes associated with cystic fibrosis due to mutation in genes in these cases. The various causes are known to occur in the prostate, the epididymis, and the testicles as well.

  • Seminal fluid analysis
    • Low volume
    • Azoospermia
    • Acid pH
    • Fructose absent

Reconstructive options for obstruction

  • Vasovasostomy – a procedure to reverse a vasectomy
  • Vasoepididyostomy or epididymovasostomy –  a surgery to reverse vasectomies
  • Transurethral incision of ejaculatory duct or TURED – surgical treatment for unblocking the ejaculatory duct obstruction

Vasectomy reversal is also associated with azoospermia, and it is one of the most common causes. It takes 3 hours to perform it and extraction of sperm is simultaneously executed to freeze as the surgery might not work.

Indications for sperm retrieval

Another reason why sperm is collected is the impossibility of some men to ejaculate for various reasons as psychological issues or retrograde ejaculation, as well as obstructive causes and so forth.

  • ejaculatory failure
  • testicular failure
  • obstructive azoospermia
    • vasa aplasia
    • vasal obstruction
    • intratesticular obstruction
    • ejaculatory duct obstruction
    • epididymal obstruction

Sperm extraction is mentioned to be quite common in IVF treatment. In this way, sperm is extracted directly from the testicles or from a sperm tube at the back of the epididymis, which is known as PESA or percutaneous epididymal sperm aspiration performed on patients with a blockage. In comparison, the procedure MESA, unlike the needles needed in the former procedure, is performed directly with an incision in the epididymis and extract sperm.

Perhaps, the more common reason why sperm extraction is performed is because some men have non-obstructive azoospermia, meaning they usually have slightly smaller testes, and their hormone profile is raised. In such cases, a different procedure is used, called MicroTESE (Microdissection testicular sperm extraction).


Patients with non-obstructive azoospermia who are encouraged to have an extraction of sperm undergo a series of examinations and investigations of genetical and hormonal checks as well as scanning and imaging of testicles to increase the chances of conception. This also includes:

  • Age of female
  • Type of sperm retrieval
  • Hormonal manipulation before SSR (Successful sperm retrieval)
  • Varicocelectomy before SRR (Successful sperm retrieval)

Another aspect that is checked is Y deletion, wipeouts on the male chromosome which do code for sperm production. If there are wipeouts present in certain regions A, B, and C, and there is a wipeout in A, B region, no sperm will be found when performing these exploratory procedures to extract or collect sperm. With C deletions, it will be possible, but that deletion will be transmitted to a male child by ICSI because it’s carried on the Y chromosome. Therefore, it’s important to do genetic testing in patients.

In theory and it is still unknown but in about 50% of men who have non-obstructive azoospermia, it’s possible to find sperm in the testicle. Several studies show that the MicroTESE technique is more precise compared to other methods and is associated with less risk, therefore this procedure is used most often around the world nowadays.

Preoperative optimization

It is said to include a series of genetics testing and counseling as well as hormonal profiles, varicocelectomy, preoperative stimulation, and ultrasound scanning before actually heading to the surgical technique.

- Questions and Answers

Have you seen any changes in male patients and sperm parameters who had gone through COVID?

From the point of view of COVID-19, we know that potentially it could affect your testes. I’ve certainly had some patients who have had COVID-19 and then develop testicular problems now we don’t know the full association. However, if you look at studies that have been done in hospitalized patients, there is quite a lot of testicular damage, and undoubtedly, we think it does probably affect it. The studies that were recently done showed about 90% of patients had very low sperm counts or no sperm after they’d had COVID-19, but again this wasn’t very well controlled in the studies. There are many other factors the patient’s been in the hospital also had various drugs, and we don’t know whether it was other effects or whether it was a COVID-19 itself that was causing uh the fertility problem.

If sperm sample is within normal range, would testing of FSH/ LH be recommended or any other testing?

No, I think if the sperm sample analysis is normal, and you’re dealing with infertility, that’s the kind of unexplained group we’re talking about. We wouldn’t necessarily do FSH and LH tested in that group as we thought it was a hormone problem, but that’s usually if the sperm count is abnormal or there’s no sperm. We’ve got to remember that a third of patients are of unknown fertility, you can have normal semen analysis and still cannot conceive. The question is, why is that? I think more and more we realize that it’s DNA damage. About a third of these patients may have DNA damage, so it’s not just about what sperm looks like on a semen analysis. It’s also about sperm integrity in terms of DNA integrity which has become much more of a novel tool that we use now in testing because there are factors that cause DNA damage. 

How does sperm DNA fragment or why?

DNA is like a ladder, and if you get a break of that ladder normally, the egg we think can repair the DNA fragmentation or damage it. As men get older, the DNA damage increases as when we get older, the egg is less able to repair the DNA, that’s the theory. There is evidence within the literature, so there tend to be environmental factors, infections, varicoceles, dietary factors, the imbalance between oxidants and antioxidants, but again the evidence for all of this is a little unclear.

One of the issues is that overall what you’ve got to look at are factors called sperm DNA damage. There may be environmental factors, we know that it may be a factor involved in embryo formation and also miscarriage,  implantation failure in patients who are trying to have IVF treatments. I co-chair a group called the European Association Guidelines Committee, we have put together some guidelines as to when we should measure sperm DNA and how we should go about that. In those guidelines, we’ve suggested that for patients who have unexplained fertility with normal semen analysis or those that might have failed IVF treatments, doing DNA fragmentation would be worthwhile testing and looking at.

You have mentioned there are 5% of patient with the trouble of having quality sperm after freezing – can you please elaborate on that? Why is this happening, and if we can know in advance that we are in this group?

I’m talking in the context of freezing sperm before operations. I always say to patients when I see them, and they come to me for sperm retrieval that we’ve done studies, others have done studies, and they showed no difference of using fresh or frozen sperm taken from the testicle or epididymis. The 5% represents all patients in those groups where we found that the sperm just did not survive the thawing process. We think that’s probably a cryopreservation process invariably, however it’s because of poor quality sperm that is frozen.

The problem is that when you face surgery on someone who has no semen or has obstructive age sperm, you’ve got to take what you’ve got, so what we try and do in those circumstances is to try and circumnavigate that is that if we find sperm on one side in operation it’s not very good we stop, and we hope that would be good enough for ICSI treatment, but then we leave the other testicle as a backup for the future to perhaps using fresh on the day.

Overall, 5% is very low, you could argue should every single patient have a synchronous cycle of ICSI with a sperm retrieval that logistically is a bit of a nightmare, it’s very difficult to arrange. That’s the kind of reason why can you mitigate that? Not really, that’s the problem, and that’s just the issue of the cryopreservation but also freezing poor quality sperm in the first place.

Did I hear that correctly the ovum can correct sperm DNA fragmentation? I’d love to know the mechanism?

That’s the theory. Some studies suggest that in theory, the younger the patient, the better ability of the egg or ovum to correct the DNA damage or repair it. That’s one of the theories that are some research is going to be done. Nobody quite knows, although we kind of look at DNA fragmentation as this kind of normal biomarker, we also need to remember the female partner’s age, the quality of her eggs as well because we so often ignore that, and our focus tends to then go to the sperm and focus a lot on the sperm.  I think we’ve got to be a little careful, but that’s a theory, and there is some evidence back in that all.

You talked about COVID, any thoughts on a vaccine before IVF concerning sperm?

 I don’t think anybody knows the answer to that, and I think that in terms of COVID-19, we know it seems to affect the testis. Giving the vaccination before in the context of prevention or transmission, the evidence for trans generational transmission, in other words, to an embryo fetus, is very extremely limited and isn’t there.  I think it’s too early to say this, but I can tell you many studies are done about this at the moment. What’s interesting about vaccination not for COVID-19, but Human papillomavirus (HPV), that’s the wart virus, is that there are thoughts about whether vaccination reduces HPV expression, which might affect IVF outcome. There is evidence, of course, that coving can affect the testis, but again the evidence is quite controversial and needs to be more robust.

When I see patients, I always ask them, have you had COVID-19?  I’ve seen patients who’ve got normal semen analysis previously because they’ve had kids, and then somebody comes up with very severe abnormalities, and I so ask them, have you had COVID-19 because I’ve seen patients who’ve certainly had been badly affected. The question is, of course, how long do you wait? Do you wait for six months and then repeat a semen analysis?  I’ve had patients who had no sperm, who’ve had COVID-19, potentially had problems in the testis and then repeated the semen analysis of 6 months, and that’s what I’m waiting for, at the moment, because it’s too early to say.

Wouldn’t you want to freeze semen anyway to prevent repeating surgical retrieval?

I was only talking in the context of freezing sperm because of an isolated procedure. If you think about a patient who doesn’t have any sperm and ejaculate and doesn’t know if you’re going to get sperm from a testicle, there are two things you can either do. One is we’re either going to do an operation on your partner, we’re going to try and extract sperm, we find it,  we’re going to freeze it. If we don’t find it, then, of course, you have an answer. 

The alternative is saying to them, look, you could do a synchronous cycle of ICSI, and we extract the eggs from the female partner, the male partner has surgery on the same day. The problem is we know that in 50% of non-obstructive age of sperm, we won’t find sperm, so what that 50% of patients who’ve gone through a cycle of stimulation should do? Do they freeze the eggs? Do they have a donor backup? The context in which I was talking about freezing sperm is yes, once we found sperm on a fresh cycle, we want to freeze sperm, and on a frozen cycle, we would freeze sperm anyway, so we would freeze what we’ve got to use.

We have morphology issues. 1% of good quality. Will this be the same on every cycle?

That depends. The problem is abnormal forms, it’s a controversial, difficult area with a huge amount of variance in reporting. You go to one lab, and they report no normal forms, you go to another lab, and they report normal forms, which is about 4%, and it is upsetting because you’re left in a situation where other parameters are normal. The question is, what do you do with these isolated abnormal forms, is it significant? Some drugs can cause abnormal forms, of course, there are congenital problems, such as globozoospermia, but as in isolation, you would normally expect all the other parameters to be abnormal if it was significant.

If you think about it, a sperm may look normal to one person, abnormal to another, strictly speaking, it shouldn’t be like that because it’s done with strict criteria called Kruger criteria, but it does happen. How do you report this? Take the abnormal forms out of the equation as a sort of factor that is confounding. You can look at the total motile count, which is the volume multiplied by concentration, multiplied by the motility percentage to get the total motile count. We think it’s probably a better indicator of fertility potential in patients.

The probability is that you don’t know whether somebody with 1% of normal forms is going to be fertile or non-fertile. The key about all of this is that the patient’s been trying. Remember, the definition of infertility is the inability to conceive after one year of unprotected intercourse. 75% of couples can conceive in a year, 95% in two years. If you do a semen analysis, you open up a can of worms in a way because if you do it pre-marriage, or you do it in 3 months in, and you find you’ve only got 1% per cent of normal forms that can have a huge psychological toll on both of you.  Does that necessarily mean that you’re infertile? No, and so I think you’ve got to be a little careful about over-interpretation of what we term as isolated teratozoospermia.

What causes slow or lack of liquefaction, and is this a concern?

Liquefaction is thought to be allowing the sperm to confer the motility of movement. Liquefaction is normal, it’s not something that I particularly have seen, if I’m honest, in many years of doing this. I think one of the issues is that liquefaction of sperm occurs due to an enzyme called PSA, which is produced by a Prostate-specific antigen, which is a mark for prostate cancer, and so it’s a protease or an enzyme that liquefies the ejaculated. That’s the process by which it occurs, if you stand in an ejaculate, it will liquefy, and you see it become runny effectively.

Its effects in terms of fertility are largely unknown in many ways, and a lot more work needs to be done in that area. Is it a concern? I would not see it as a major concern, because effectively, we don’t know what the role of liquefaction is in terms of sperm function, but we think it is involved in the sperm’s motility in terms of the female partner when it’s in the female genital tract.

Can you tell me more about gonadotropins? Do they work?

I think you’re referring to hormone stimulation in the context of having azoospermia or having no sperm or hormone therapy. What can be done in some men, people argue, but gonadotrophins, which is FSH and LH stimulation to raise testosterone potentially, stimulate sperm production in that way. The other way of doing it is with anti-estrogens to stimulate FSH/LH gonadotropins to stimulate testosterone and sperm production.  The evidence for this is quite limited, the problem is when people do trials, they use their patients themselves, and as I said, some patients produce sperm, and that can appear and disappear. Do they work? We don’t know that, and we’re doing a study at the moment, but because of the confidentiality reasons tell you the result of that, at the moment, it suffices to say that the data is quite mixed and very difficult to interpret. We’ve done a meta-analysis hopefully, in about a month, we’ll be able to share that data.

The only time I would be giving it anyway is to stimulate testosterone in the men who have low testosterone because then you’ve got kind of nothing to lose because remember these kinds of drugs are expensive and not without side effects. In fact, in men, they’re off label, we’ve got to remember they’re not licensed to be used in men, they’re licensed in women, so that’s where the difficulty and controversy about all of this come from.

Some clinics use ICSI as a standard. When there is no known reason for infertility, is there any benefit to doing this, or can the risk of the process be greater than any benefits?

If you look at results from ICSI, they seem to be superior to IVF treatments, and that’s the kind of argument that you know about that people use. The evidence for IVF treatments causing congenital problems is limited, I’m not an IVF doctor as such, but ICSI can be associated with other congenital problems, however, it’s weighing up results and what’s best, and that’s where the sperm count comes in that’s what the sperm is.  Ideally, ICSI should be used for male factor infertility, and more and more, we tend to use these type of treatments as well as IMSI and other treatments for DNA fragmentation. It depends on the IVF unit, I suppose in many ways, this doesn’t seem to be any standardization of this, and that’s one of the problems, and I think that’s what needs to be addressed by the HFEA, so perhaps we are overusing ICSI in that setting. 

For surgically retrieved sperm, what is the conventional method to freeze? “Neat” with tissue debris, or processed in some way?

Both, so they can extract the sperm and that way alternatively sometimes when they’ve got tissues still they can freeze the tissue. I’m not an embryologist, but that would be a more embryological question, but they could do it both ways.

What would you suggest if there is 60% of dead sperm? What could cause this, and is it still possible to conceive naturally?

In a sample, you will get dead sperm, that’s the natural cycle, so what causes non-vital sperm, in other words, sperm that’s not alive. In that context, you need to look at infections. I’ve seen that urinary tract infections Sexually transmitted infections (STIs).

You would normally see either infection, but if you have a congenital problem, you can sometimes get various disorders that can get immotile sperm. Some of these conditions are called Kartagener’s or Immotile cilia syndrome, which is associated with chest problems as well where the flagella or tails aren’t moving, and the sperm, in a sense, is alive but depends on how you define vitality it’s a general vital stain on the cells. The most common one we see is, in terms of infection, but again the thresholds that we use are controversial, various drug treatments potentially gonadotoxic or toxins to the testis can also cause this, but you would have normally seen very severe cases of this. If I see non-vital sperm, one thing I would be looking at would be how many white cells are in the ejaculate, white cells might indicate infection as well. I’d probably want to culture the sperm and also do some other infection tests to make sure there isn’t an ongoing infection causing them. 

Can DNA fragmentation testing be carried out on sperm collected by Micro TESE, if yes, does it matter if the sperm is fresh or frozen?

It is only a research setting that it’s done currently, but it can be done and the issue about Micro TESE is the question you need to ask yourself: why are you doing Micro TESE in the first place in this setting. There are two reasons why you do Micro TESE.

First is non-obstructive azoospermia, which obtains sperm, and then you freeze it and use that sperm for ICSI treatment. The other setting where you do testicular sperm extraction for men who are non-azoospermic, who have sperm and ejaculate is as a kind of treatment in patients who have raised DNA fragmentation or damage, where you want to try and bypass the DNA fragmentation. The argument being the testicular sperm in some studies have shown to have less fragmented DNA than ejaculated sperm. That’s been shown in some studies, so you can measure DNA fragmentation within testicular sperm and compare it and ejaculate it. The other context we sometimes can argue using would be in patients who might have sperm frozen, and they’ve had multiple cycles because there is an argument that freezing sperm can damage your DNA. They might want to do it to see whether they’re failing embryogenesis or failing IVF cycles using frozen sperm and if it is due to the DNA damage to the sperm. The problem is that its patients who have these kinds of problems, it’s like gold dust because again you’ve done an operation to extract that sperm, therefore it’s a test that you’ve got to be very careful about doing. 

To answer the question directly, you can measure DNA damages in testicular sperm, you would normally do a testicular sperm extraction on the day of an ICSI cycle as a method to try and bypass or try and circumnavigate, mitigate the problem of sperm ejaculated sperm and its effect in terms of DNA damage. The argument being that testicular sperm has less fragmented DNA. You’ve got to be cautious about this because levels of evidence for this are extremely limited. We’ve written guidance on this and said that if you’re going to be doing this, ideally, it should be done in a research setting. 

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Professor Suks Minhas

Professor Suks Minhas

Professor Suks Minhas is a consultant Urologist and lead Andrologist at Imperial College, London. He qualified from the University of Sheffield. He did his post-fellowship training in andrology at University College London. He obtained a research degree from the University of Leeds which was awarded a commendation. His expertise is Andrology, male infertility, testicular cancer, Peyronie's disease, microdissection TESE, penile reconstruction, penile implants, erectile dysfunction, testis pain, sexual dysfunction in men, penile problems, treatments for male fertility He is an expert in male fertility and men's health including disorders of the male genital system and testis cancer. A recipient of the prestigious Karl Storz Telescope, an award given to specialists who have made a significant and lasting contribution to British urology, he is recognized as a key international opinion leader in his field. Professor Minhas specialises in treating problems affecting the male genitals and is one of the UK’s only dedicated andrological surgeons. In addition, he has published extensively on the subject, with 195 peer-reviewed papers, including a recent textbook on male sexual dysfunction. He has chaired over 80 scientific sessions at meetings both nationally and internationally and has given almost 300 invited lectures internationally. Professor Suks is also a referee to a number of scientific journals and a member of international guideline committees including the EAU guidelines on male sexual and reproductive health (2020). He is actively involved in the teaching and training of surgeons in the UK and co-ordinates fellowship training programmes. He is a board member of the European Society of Andrological Urology (ESAU). He is a firm believer in evidence-based practise and is co-chair of the European Association of Urology guidelines on male sexual and reproductive health. He was chairman of the British Association of Urological Surgeons Section of Andrology and is a faculty member of the European School of Urology.
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