Assisted Embryo Hatching
• Assisted hatching is an IVF technique in which the zona
is treated prior to embryo transfer in order to weaken
the wall of the embryo and thus improve the likelihood
of successful hatching and embryo implantation.
• Just prior to embryo transfer, the developing embryo
must"hatch” out of its outer shell (zona pellucida).
• Some embryos seem to have a thicker shell that may
decrease their ability to implant
• In such cases Advance technique of IVF with assisted
hatching is performed.
Indications for Assisted Hatching:-
•Elevated maternal age
•Elevated follicle stimulating hormone [FSH] level
•Evidence of a thickened zone pellucida of the embryos in IVF
•Unexplained implantation failure after two or more embryo
•Frozen thowed embryo.
•Excessive fragmentation >20%
Embryo hatching with laser Blastocyst
hatching in preparation human embryo at
Advantages of assisted embryo hatching
Assisted hatching has demonstrated the potential for improving
embryo implantation rates and clinical pregnancy rates in select
patient groups undergoing IVF.
- Fewer embryos required for transfer.
- Increased implantation success rates
- Allow for blastocyst culture.
Techniques of Assisted Embryo Hatching
• Mechanical Hatching
• Chemical Hatching
• Laser Hatching
• In this the embryo cover is slit open mechanically with the help of
a thin long drawn out glass needle.
• This is done with the help of a machine called micromanipulator
• The zona pellucida is pierced with a very thin glass micro
needle through both sides, the needle tip being controlled in the
perivilelline space by eye.
• Then the suction of the holding pipette is stopped and the holding
pipette is rubbed against the trapped area of the zona until this area
has been completely abraded.
• Creating a hole in the zona pellucida of an embryo by using
acidic tyrode’s solution .
• The fine micropipette containing acidic tyrode’s solution is
brought very close to the zona pellucida and the acidic solution
expelled gently over a small area (~20-30mm) until the zona is
dissolved through to the inside.
• After the procedure the embryos are thoroughly washed in fresh
medium and cultured until the time of transfer.
Acidic tyrode’s solution of pronase is used for
drilling holes in zona pellucida.
Laser assisted Hatching
• This technique involves the creation of a precise gap in the zona
pellucida of selected embryos using a 1.48 micron infrared diode
Contact laser / Non contact laser
• Contact laser: laser radiation has to be delivered directly to the
zona called contact laser
• Non contact laser: laser gamete manipulation has described in the
non-contact mode. Using various wavelengths.
• It allows direct delivery of laser beam through the objectives.
• 1.48 micron diode laser is advantageous over all other
techniques as at 1.48 wave length , radiation is non mutagenic
• The radiation is administered in the non contact mode without
Assisted hatching done by using laser light
The embryo is held
with gentle suction
using a glass pipette
The red “pilot light “
provides a visual target
for the laser.
The white circle
marks the safety region
based on temperature.
The laser is usually
fired 3 times to
produce a small hole in
Importance of Laser Assisted Hatching
• This specifically designed laser system includes the laser,
which serves as the energy source to create an opening in the
• A computer which allows the operator to precisely control the
laser energy output, laser pulse duration and gap size.
• Takes less time and does not expose the embryos to potentially
• This procedure appears to be quick, precise and is chemical
If required this facility
Please contact us:
Dr. Rama's Institute for Fertility
6-3-788/33, Opp. Chandana Brothers,Ameerpet,
Hyderabad - 16, Andhra Pradesh, India.
Phones: +91-040- 23401135, 23402294.