IVF Process Step by Step at 6Venus Fertility Hospital, Wakad Pune

More than 12 million babies have been born worldwide through IVF since 1978. In India alone, over 2.5 lakh IVF cycles are performed every year – and that number keeps climbing. Despite this, most couples walking into their first fertility consultation have a very incomplete picture of what the IVF process step by step actually involves. They know it has injections. They know it has a lab. Beyond that, the process feels abstract and intimidating.

At 6Venus Fertility and Urology Hospital in Wakad, Pune, Dr. Pavan Bendale walks every couple through each stage before treatment begins – not to fill time, but because couples who understand what is happening and why are better prepared, more compliant with protocols, and carry significantly less anxiety through the cycle. Clarity is part of clinical care here.

What follows is the complete IVF procedure steps as they happen at 6Venus – with the clinical detail that most online guides leave out.

Before the Cycle Begins: The Diagnostic Foundation That Decides Everything

The most consequential part of the IVF process step by step is not the egg retrieval or the embryo transfer. It is what happens before a single injection is given. No two IVF protocols should be identical, because no two patients are identical. The diagnostic phase is where that personalisation begins.

At 6Venus, both partners are evaluated simultaneously from day one. This matters because male factor infertility contributes to around 30-40% of all infertility cases, yet many clinics delay male evaluation by weeks. Losing that time costs cycles.

Female partner baseline tests include:

• AMH (Anti-Mullerian Hormone) – the best indicator of ovarian reserve
• FSH, LH, Estradiol on Day 2 or 3 of the menstrual cycle
• Thyroid profile (TSH, T3, T4) – hypothyroidism silently disrupts ovulation and implantation
• Prolactin – elevated prolactin suppresses ovulation and IVF response
• Transvaginal ultrasound – antral follicle count, uterine structure, ovarian cysts
• Infectious disease screening – HIV, Hepatitis B, Hepatitis C, VDRL
• Hysteroscopy (where indicated) – to rule out polyps, adhesions, uterine septum

Male partner baseline tests include:

• Semen analysis – count, motility, morphology (Kruger strict criteria)
• Sperm DNA fragmentation index (DFI) – where lifestyle risk factors or prior failure indicate it
• Infectious disease screening – matching female partner panel
• Hormonal evaluation – if sperm parameters are severely abnormal

Dr. Bendale reviews these results in detail before any protocol is designed. The AMH level determines stimulation dose. The antral follicle count confirms reserve. The semen parameters decide whether standard IVF or ICSI is more appropriate. Skipping this foundation and jumping to injections is clinical guesswork. For a deeper understanding of male fertility evaluation before IVF, see the 6Venus guide on male fertility tests before IVF or IUI.

Step 1 – Protocol Selection: The Decision That Shapes Your Entire Cycle

Once test results are reviewed, Dr. Bendale selects a stimulation protocol. This is a clinical decision with real consequences for how many eggs are retrieved, how the ovaries respond, and whether the cycle proceeds to a fresh transfer or a freeze-all. There is no single correct protocol – the choice depends on your specific ovarian reserve, age, and any previous cycle history.

The main protocols used at 6Venus include:

• Long agonist protocol: Begins mid-luteal phase with a GnRH agonist to suppress natural hormones before stimulation starts. Preferred for patients with good ovarian reserve. Gives the doctor precise control over the stimulation window.
• Antagonist protocol: Stimulation begins on Day 2-3 of the cycle with gonadotropins, and a GnRH antagonist is added mid-stimulation to prevent premature LH surge. Shorter, more flexible, preferred for poor responders and PCOS patients due to lower OHSS risk.
• Mini or mild IVF: Lower-dose stimulation for women with very poor reserve or for those who have responded badly to standard doses previously. Retrieves fewer eggs but reduces physical and financial burden per cycle.
• Natural cycle IVF: No stimulation – works with the body’s single naturally maturing egg. Used in select cases of very poor reserve where ovaries do not respond to stimulation at all.

The protocol is not a formality. An incorrect protocol for a PCOS patient can trigger dangerous ovarian hyperstimulation syndrome (OHSS). An insufficient protocol for a low-responder wastes a cycle. Getting it right at the start is one of the most important contributions an experienced IVF doctor makes.

Step 2 – Ovarian Stimulation: 8-12 Days That Grow Your Eggs

Stimulation begins on Day 2 or Day 3 of the menstrual cycle. The female partner self-administers daily subcutaneous injections – into the lower abdomen – containing gonadotropins (FSH and sometimes LH) that signal the ovaries to mature multiple follicles simultaneously. In a natural cycle, one follicle matures and releases one egg. IVF stimulation encourages several follicles to develop at once, increasing the number of eggs available for retrieval.

The injections themselves are done with very fine needles and take under a minute. Most women manage this comfortably after the first one or two attempts, especially once the 6Venus nursing team demonstrates the technique and answers questions in person.

What to expect physically during stimulation:

• Mild bloating and pelvic heaviness as follicles grow
• Breast tenderness and mild mood fluctuations from rising estrogen
• Occasional fatigue – rest when needed
• No heavy exercise, alcohol, or smoking during this phase
• Increased fluid intake – important for ovarian health and OHSS prevention

Stimulation medications are billed separately from the procedure package and represent the largest variable in total IVF cost. The dose depends entirely on your ovarian reserve and response. For a full breakdown of what each stage of treatment costs at 6Venus, see the IVF treatment cost guide for Wakad patients.

For detailed guidance on how diet during this phase can support your stimulation response, see the 6Venus article on fertility diet planning in Pune.

Step 3 – Monitoring Scans: What the Doctor Is Watching and Why

Every 2-3 days during stimulation, the female partner attends a monitoring visit at 6Venus. These visits involve a transvaginal ultrasound scan and sometimes a blood test for estradiol levels. The scan checks:

• Follicle count – how many follicles are developing in each ovary
• Follicle size – follicles need to reach 16-20mm before retrieval; too small means the egg inside is immature
• Estradiol level – rising estradiol confirms follicle growth; an unusually rapid rise flags OHSS risk
• Endometrial thickness – lining should be reaching 7-10mm with a trilaminar (triple-layer) pattern by retrieval day for a fresh transfer

For couples based in Wakad, Hinjewadi, and Pimpri-Chinchwad, 6Venus’s location on Dange Chowk Road means these monitoring visits can be completed early morning without disrupting work schedules. This practical reality matters across a 10-12 day stimulation phase with multiple visits.

Step 4 – The Trigger Injection: Precise Timing Is Everything

When the lead follicles reach 18-20mm and estradiol levels confirm readiness, Dr. Bendale gives the trigger shot. This is a single injection of hCG (human chorionic gonadotropin) or a GnRH agonist trigger, administered at a very precise time – usually between 10 PM and midnight – because egg retrieval must happen exactly 34-36 hours later. Giving the trigger even a few hours early or late shifts the eggs’ maturity window and reduces retrieval quality.

The trigger injection is the most time-sensitive part of the IVF procedure steps. The 6Venus team gives couples a specific administration window with the exact time written out, and confirms it is followed. Do not miss this injection and do not administer it at the wrong time.

Step 5 – Egg Retrieval: The Procedure Itself

Egg retrieval – also called ovum pick-up (OPU) – happens 34-36 hours after the trigger injection. It is a day procedure. No hospital admission is needed. The patient arrives fasting, is given light intravenous sedation (not general anaesthesia), and the procedure takes 15-25 minutes.

Dr. Bendale guides a fine transvaginal needle into each ovary under ultrasound guidance and aspirates the follicular fluid from each follicle. That fluid contains the eggs. The embryology team receives the fluid immediately and isolates mature eggs under the microscope in the adjacent laboratory.

What happens after retrieval:

• Recovery in clinic: 1-2 hours of observation before discharge
• Mild cramping and spotting is normal for 24-48 hours
• Rest that day; most patients return to desk work the next day
• The embryology team calls with the egg count the same afternoon
• A second call follows the next morning with the fertilisation result

On the same day as retrieval, the male partner provides a fresh semen sample at the clinic. If the male partner has azoospermia or very poor sperm quality, surgical sperm retrieval (TESA or Micro-TESE) is coordinated to coincide with egg retrieval day. 6Venus performs both procedures in-house, which eliminates the coordination delays that arise when sperm retrieval is sent to an outside urology centre. More on this is covered in the 6Venus article on TESA and Micro-TESE advanced sperm retrieval.

Step 6 – Fertilisation: IVF vs ICSI – Which Happens in Your Cycle

Within 2-4 hours of retrieval, the isolated eggs and prepared sperm meet in the 6Venus embryology laboratory. How they are combined depends on your diagnosis.

Standard IVF insemination:

50,000 to 100,000 prepared sperm are placed alongside each mature egg in a culture dish. Fertilisation happens naturally if the sperm penetrates the egg. Used when semen parameters are normal. Results are checked 16-18 hours later – fertilised eggs show two pronuclei (2PN), confirming successful fertilisation.

ICSI (Intracytoplasmic Sperm Injection):

A single selected sperm is injected directly into each mature egg using a glass micropipette under high magnification. Used when sperm count, motility, or morphology is abnormal; when sperm was surgically retrieved; when previous IVF had poor or zero fertilisation; or when very few eggs were retrieved and maximum fertilisation efficiency is needed. On average, 70-80% of mature eggs fertilise successfully with ICSI.

The decision between IVF and ICSI is made before the cycle based on the semen analysis. Sometimes Dr. Bendale recommends ICSI for a cycle where the semen parameters look borderline – to protect against unexpected fertilisation failure. For a clear comparison of when IVF versus other treatment paths apply, see the 6Venus breakdown of IVF vs IUI.

Step 7 – Embryo Culture: What Happens in the Lab Over 5 Days

After fertilisation is confirmed, embryos are placed in a precisely controlled incubator environment – the most critical piece of infrastructure in any IVF laboratory. Temperature is held at exactly 37°C. Oxygen and CO2 levels are monitored continuously. pH is maintained within a narrow range. Any deviation from these parameters stresses developing embryos and reduces quality. This is why in-house embryology – run by a dedicated team at 6Venus rather than outsourced to a shared lab – matters for outcomes.

The developmental stages embryologists watch:

• Day 1: Fertilisation check – 2 pronuclei visible confirms successful fusion
• Day 2-3: Cleavage stage – embryo divides into 4-8 cells; graded on cell symmetry and fragmentation
• Day 4: Morula – embryo compacts into a mulberry-shaped cell mass
• Day 5-6: Blastocyst – embryo develops a fluid-filled cavity (blastocoele) with distinct inner cell mass and trophectoderm; only the strongest embryos reach this stage

Not all embryos that fertilise will reach blastocyst stage. On average, 40-60% of fertilised eggs develop to blastocyst. This attrition is natural and is part of biological selection – it happens in natural conception too, invisibly. The embryos that do not reach Day 5 likely had chromosomal issues that would have prevented successful implantation anyway.

Blastocyst transfer is preferred at 6Venus when the number of developing embryos supports it, because a blastocyst that has survived to Day 5 has already demonstrated developmental competence. Transferring a Day 3 embryo is a less informed bet. For a full explanation of how embryo grading affects your cycle, see the 6Venus resource on embryo quality and what it means for IVF success.

Step 8 – Pre-implantation Genetic Testing (PGT): The Optional Step That Changes Outcomes

For couples who qualify – women over 37, those with recurrent pregnancy loss, previous failed IVF cycles, or a known genetic condition – PGT is performed at the blastocyst stage before transfer. A small biopsy of 5-8 cells is taken from the trophectoderm (the outer layer that forms the placenta) and sent to a genetics laboratory for chromosomal analysis.

Only embryos confirmed as euploid (correct chromosomal number, 46 chromosomes) are transferred. This step adds 1-2 weeks to the cycle timeline because results take time to return. During this period, all biopsied blastocysts are vitrified (fast-frozen) and stored. The transfer then occurs in a subsequent frozen embryo transfer cycle.

Who benefits most from PGT:

• Women 37 and older – chromosomal abnormality rate in embryos rises sharply with age
• Couples with two or more miscarriages – chromosomally abnormal embryos are the leading cause
• Couples with two or more failed IVF cycles despite good-quality embryos
• Carriers of known genetic conditions (PGT-M) or chromosomal rearrangements (PGT-SR)

According to published data from the National ART Registry of India (NARI) reviewed on PubMed, ICSI is performed in over 52% of Indian IVF cycles due to male factor infertility, while PGT is increasingly adopted as a standard of care in specialised centres. At 6Venus, Dr. Bendale was among the early adopters of PGT in the Wakad-Pune region.

Step 9 – Embryo Transfer: The Simplest Procedure With the Highest Stakes

Embryo transfer is the gentlest step in the entire IVF process step by step. No anaesthesia. No sedation. The procedure takes under 10 minutes. The female partner arrives with a comfortably full bladder (which straightens the uterine angle for easier catheter passage) and lies on the procedure table while Dr. Bendale passes a soft, flexible catheter through the cervix under ultrasound guidance, depositing the embryo into the uterine cavity at the optimal position.

At 6Venus, a single embryo transfer (SET) is the standard approach for most patients – particularly women under 38 with good-quality blastocysts. Transferring two embryos does modestly increase pregnancy odds but significantly raises the risk of twins, which carries higher complications for both mother and babies. Dr. Bendale discusses the embryo number decision openly with each couple based on age, embryo quality, and history.

Fresh vs Frozen Embryo Transfer (FET) – an important distinction:

Factor	Fresh Transfer	Frozen Embryo Transfer (FET)
Timing	Same cycle as retrieval (Day 3 or Day 5)	Subsequent cycle – 4-6 weeks later
Uterine environment	May be suboptimal if estrogen/progesterone elevated after stimulation	Cleaner – uterus fully prepared separately
Success rates	Good in most patients	Equal or higher in many patient groups
OHSS risk	Higher if ovaries were very responsive	No OHSS risk – stimulation already completed
When preferred	Normal responders with good lining	PCOS, high responders, PGT cycles, thin lining

When a freeze-all cycle is recommended – which happens when OHSS risk is elevated, lining is suboptimal, or PGT is being done – all embryos are vitrified and transfer occurs in a planned frozen cycle. The success rates of a well-executed FET are at least as good as, and sometimes better than, a fresh transfer in the same cycle. For more detail on what happens when an embryo does not implant, the 6Venus resource on embryo implantation failure covers this in clinical depth.

Step 10 – The Two-Week Wait and the Pregnancy Test

After embryo transfer, the female partner takes progesterone supplementation – via vaginal pessaries, injections, or both – to support the uterine lining and sustain early implantation. This continues until the pregnancy test and, if positive, for several more weeks into early pregnancy.

The wait between transfer and the pregnancy blood test is typically 10-14 days. It is the most emotionally demanding stretch of the entire cycle. Physical symptoms during this period – cramping, spotting, bloating, breast tenderness – do not reliably predict outcome. These symptoms can occur whether the cycle has succeeded or not, because progesterone supplementation itself causes them.

The 6Venus team is reachable during this period for questions. Dr. Bendale’s approach is to discourage home pregnancy testing before the scheduled blood test date – home tests can give false negatives in early implantation and false positives from the hCG in the trigger shot if tested too early, both of which cause unnecessary distress.

The pregnancy blood test measures beta-hCG – a hormone produced by implanting embryos. A positive result leads to a scan 10-14 days later to confirm a heartbeat. A negative result triggers a review consultation where Dr. Bendale goes through the cycle in detail to understand what can be changed before the next attempt.

What Happens After IVF – Frozen Embryos, Failed Cycles, and Next Steps

If good-quality embryos remain after transfer, they are vitrified using the glass-freezing technique and stored at 6Venus. Vitrification has transformed embryo storage – survival rates after thaw exceed 95% at specialist centres, and frozen embryo transfers now achieve results comparable to fresh cycles. These stored embryos are your resource for subsequent FET cycles without repeating the stimulation and retrieval phase.

If the first IVF cycle does not result in a pregnancy, the conversation at 6Venus does not start with “let’s try the same thing again.” It starts with a systematic review of what happened at each stage – how many eggs, how many fertilised, how embryos developed, what the lining looked like, and whether the transfer was technically smooth. That review shapes the adjusted protocol for the next attempt.

Recurrent failure – defined as failure after two or more good-quality embryo transfers – triggers a deeper investigation: hysteroscopy if not already done, sperm DNA fragmentation testing, immunological and thrombophilia screening, and often PGT if embryo quality was the suspected issue. The 6Venus clinical position on why embryos fail to implant is covered in full at the embryo implantation failure page.

IVF at 6Venus Wakad – The Integrated Advantage

Most fertility clinics handle female infertility. 6Venus handles both, under one roof. Dr. Pavan Bendale leads the fertility wing. Dr. Sunil Palve, MCh Urology, leads the urology wing. When the male partner’s infertility has a urological root – varicocele, obstructive azoospermia, hormonal disruption – both specialists work on the same case simultaneously. No referrals. No delays. No reports getting lost between providers.

For couples in Wakad, Hinjewadi, Pimple Saudagar, Marunji, and Pimpri-Chinchwad, the proximity of 6Venus to the Hinjewadi IT corridor makes the monitoring-intensive stimulation phase manageable around work schedules. The clinic’s in-house semen analysis and diagnostic capabilities mean baseline testing does not require multiple external lab visits before the IVF process begins.

As the NHS’s IVF overview confirms, the success and safety of an IVF cycle depend heavily on clinical judgment at every step – protocol selection, monitoring decisions, fertilisation method, embryo assessment, and transfer timing. These are not standardised algorithmic steps. They are decisions made by an experienced clinician reading a living biological process in real time.

To understand what the IVF process step by step looks like for your specific diagnosis and reports, the most useful starting point is a consultation with Dr. Pavan Bendale at 6Venus Fertility and Urology Hospital, Darekar Heights, Dange Chowk Road, Wakad, Pune – 411033.

10 Questions Couples Ask About the IVF Process Step by Step

1. How long does one IVF cycle take from start to finish?

A single IVF cycle – from the first day of stimulation to the pregnancy blood test – takes approximately 4-6 weeks. This includes 8-12 days of stimulation, the egg retrieval, 5-6 days of embryo culture, and 10-14 days after transfer before the blood test. If a frozen embryo transfer is planned in a separate cycle, add another 4-6 weeks. Pre-cycle testing and protocol planning add time before this, especially if results require follow-up investigation.

2. Are the injections during IVF painful?

Most patients describe the stimulation injections as mild discomfort rather than pain. The needles are very fine and go subcutaneously (just under the skin) in the lower abdomen. The first two or three injections are the most unfamiliar. After that, most women manage independently without difficulty. The 6Venus nursing team demonstrates the injection technique at the start of the cycle and is available by phone for any concerns during the stimulation phase.

3. How many eggs are typically retrieved in an IVF cycle?

The number depends entirely on ovarian reserve and stimulation response. Women with good reserve and normal AMH typically retrieve 8-15 eggs. Low responders may retrieve 2-5. Women with PCOS who hyper-respond may produce 20 or more follicles, though not all contain mature eggs. Of the eggs retrieved, approximately 70-80% will be mature and suitable for fertilisation. Not all will fertilise. Not all fertilised eggs will develop to blastocyst. This progressive attrition is normal and expected.

4. What is the difference between IVF and ICSI?

In standard IVF, eggs and sperm are placed together in a dish and fertilisation happens naturally. In ICSI, a single sperm is injected directly into each egg. ICSI is used when sperm parameters are abnormal, when sperm was surgically retrieved, or when previous IVF cycles had poor fertilisation. Both happen in the lab – the difference is in how egg and sperm are combined. ICSI does not guarantee better outcomes in couples with normal sperm; standard insemination is sufficient and appropriate in those cases.

5. What is a blastocyst transfer and why is it better than Day 3 transfer?

A blastocyst is an embryo that has developed for 5-6 days in the laboratory. By Day 5, only embryos with sufficient developmental potential reach blastocyst stage – making the selection more self-filtered. Transferring a blastocyst means transferring an embryo that has already passed a natural developmental checkpoint. Day 3 embryos look similar morphologically but their eventual fate is less certain. Blastocyst transfer is preferred at 6Venus wherever the number of developing embryos makes it viable. See the 6Venus resource on embryo quality for more.

6. What is a freeze-all cycle and when is it recommended?

A freeze-all cycle means all embryos are vitrified after culture and no fresh transfer is done in the stimulation cycle. Transfer happens in a separate, planned frozen embryo transfer (FET) cycle. Freeze-all is recommended when OHSS risk is high (often in PCOS patients), when the endometrial lining is not optimal on retrieval day, when PGT is being performed, or when progesterone levels rise prematurely during stimulation. FET success rates are comparable to or sometimes better than fresh transfer – the uterus gets a clean, well-prepared environment rather than one still recovering from stimulation.

7. Can the male partner’s sperm be frozen in advance for IVF?

Yes. If there is any concern about the male partner being able to produce a sample on retrieval day – due to travel, anxiety, or known intermittent sperm production – a backup frozen sample can be collected and stored in advance at 6Venus. This is a straightforward precaution that eliminates the risk of retrieval proceeding without viable sperm available. The 6Venus egg and sperm freezing service handles this as part of pre-cycle planning. See the egg and sperm freezing page for details.

8. What should the female partner avoid doing during the stimulation phase?

During stimulation, the ovaries enlarge significantly. Avoid: high-impact exercise, heavy lifting, anything that risks ovarian torsion (twisting). Alcohol and smoking are strictly off-limits. Sexual intercourse is generally advised against once follicles exceed 15mm to reduce torsion risk. Continue with normal light activity – walking is fine and encouraged. Stay well-hydrated. Maintain a protein-rich diet to support follicle development. Follow the 6Venus fertility diet plan for specific food guidance during the stimulation phase.

9. How soon after a failed IVF cycle can we try again?

If you have frozen embryos from the first cycle, a frozen embryo transfer can be planned as soon as 4-6 weeks after the failed transfer, once the uterus has returned to its natural cycle and Dr. Bendale has reviewed the case. If a new stimulation cycle is needed, most doctors recommend waiting one full natural menstrual cycle to allow the ovaries and body to recover before beginning stimulation again. The gap is also a clinical opportunity – to run additional investigations, adjust protocol, and make changes that give the next attempt a better foundation.

10. What happens to embryos that are not transferred or frozen?

Embryos that stop developing before reaching a viable stage (early arrest) are not suitable for use and are not frozen. Viable embryos that are not transferred in the current cycle are vitrified and stored for future use. At 6Venus, storage is handled in-house and couples are informed of their embryo inventory after every cycle. Annual storage fees apply after the first year. Couples who complete their family and have remaining frozen embryos discuss their options with Dr. Bendale – which may include continued storage, compassionate transfer, or other medically and ethically appropriate decisions.