Dedicated to Women
 

Baheti Hospital

&Centre for Reproductive health care

 

(Test Tube Baby Centre)

Dedicated to Women's Health
 
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IVF Lab Tour
 

1.

Any woman undergoing IVF/ICSI treatment receives hormone injections during the first 12 days of her menses cycle to make her produce at least more than 5 follicles resulting in increased chance of conceiving in that cycle.   To monitor the development of ovarian follicles, Ultrasound  examination is done 3-4 times in that cycle along with checking of estradiol level in her blood.   Well stimulated ovary will look like this under trans-vaginal ultrasound examination.

Ultrasound Picture of Ovary with multiple follicles
 

 

2.

About approximately 12-14 days after such hormone injections, patient is ready for egg collection.  On the day of egg collection, patient is given mild sedation and local anaesthesia, follicular fluid is collected with the help of an aspiration pump and a special egg collection needle under trans-vaginal ultrasound guidance, taking care to keep the temperature of follicular fluid at 37’C.

Egg Collection showing Aspiration pump, US machine

EC needle
 
 

3.

Eggs are identified with the help of a Stereo-zoom microscope, removed from

the blood-stained follicular  fluid, washed in  Culture  medium  and  kept   for

culture in a CO2 incubator.

 

Egg Identification in Progress under laminar air flow

Mature Egg

   

Eggs are handled in sterile environment under bacteria-filtered positive air pressured room under a laminar air flow module.

Laminar Air Flow

Clean Air positive pressure

Module

The culture media used for growth of embryos are mostly ready-to          use imported material obtained by us every month.   As a quality        control measure, these media are checked randomly for pH and      osmolarity using pH meter and osmometer.
 
Osmometer and Balance Petri dish covered with Oil having media droplet pH meter
   
   

Baheti Hospital’s IVF Unit is equipped with two CO2 Incubators and both

are running simultaneously as a backup support in case any one of them

fails.Both these incubators and all the other IVF equipment are getting

clean and uninterrupted power supply through online UPS. In case of

equipment failure or CO2 gas exhaustion, an external siren gets activated

to alert not only the Embryologist but all the staff of the Hospital to enable

them to attend to the faults and rescue the precious eggs/embryos

instantly. Besides the UPS, we do have the support of a generator to     give us power supply during occasional long power failure.

CO2 Incubators Uninterrupted Power Supply UPS

      Siren (Alarm) to            alert  the Staff

 
 
 
 

Once the eggs are in the incubator, husband’s semen is washed and

motile sperms are collected for further processing for either IVF or ICSI.  

For fertilization, washed sperms from the husband’s semen are added

in required number to the eggs in case of IVF.

In case, the sperms are low in number or are of poor motility grade, ICSI is

done on eggs with the help of a micromanipulator. Size of an egg is

approximately 0.15 mm and the size of a sperm is approximately 0.005 mm.

Hence, an inverted microscope is needed to magnify them to 200 to 400

times so that they can be visualized during the procedure.  To do the high-precision ICSI work,  this microscope has been improvised with a

micromanipulator to hold an egg on one side and pick up and inject a single

sperm into a single egg through the opposite side as shown below.

 

Micromanipulator ICSI
   

After IVF/ICSI, cultured embryos are examined the next day morning for any sign of fertilization which is 2 PN (one pronucleus each from husband and wife) appearing inside the egg.

Fertilized Egg 2 PN (Pronucleus)
 

If too many surplus embryos are there (for e.g. more than 12) some can be frozen-stored at this stage for future use for the patient couple.
On further culturing, 2PN embryos will divide to become either 2-cell or 4-cell by next day morning.
2-Cell Embryo 4-Cell Embryo 8-Cell Embryo
     

On further culturing, by Day 5, embryos will develop to a stage called Blastocyst

Blastocyst and a Hatching Blastocyst Embryo Transfer Catheter

Remaining surplus embryos are cryopreserved for future use for the same patient in case of failure of this fresh IVF/ICSI cycle.  Embryos are frozen in small tubes called straws (which are labeled with patients name, date) using this freezing machine and are stored under liquid nitrogen(-196’C).

 

 

Freezing equipment at Baheti Hospital Embryos Stored in Straws which are inside Canisters
 
   

Embryo Storage Container Some of our Successes
 
   
14

14, Usha Colony, Malviya Nagar Main Road, Jaipur, Rajasthan (INDIA)
Tel: 91-141-2754049, 2754408, 3256810. E-mail: bahetihospital@gmail.com