Acrosome Intactness gelatin film on the glass slide is lysed coming in contact

with proteolytic enzymes leaching out of acrosome and

test allow light to pass through creating a halo around the

sperm head which can be examined using a microscope.
GELATIN FILM LYSIS TEST KIT
Material included with the Kit
The sperm acrosome is located on the top of the sperm
head. During acrosome reaction, the outer acrosomal 1. Incubation reagent 0.5 ml x 10 ampoules
membrane and the plasma membrane fuse and vesisculate 2. Pre-coated Gelatin Film Slides 10 no.
to discharge the acrosomal contents. Multiple enzymes are
present in the sperm acrosome including several acid
hydrolases commonly found in lysosomes. Acrosomal Material not included with the Kit
enzymes help the sperm to penetrate the cumulus mass and
the zona pellucida 1. Pipettes

A sperm with defective acrosome may not have the

2. Incubator at 50oC.
3. Petri dish with moist filter paper
sufficient stock of these proteolytic enzymes and would
eventually fail to penetrate the zona, a prerequisite for
successful fertilization.
Storage and Stability :
Store below 30oC till expiry indicated on label.
Sperm acrosome status and function may be assessed by
various methods including the measurement of direct
proteolytic activity present in the acrosome.

Gelatin film lysis is one such method which detects

acrosome proteolytic activity in terms of gelatin digestion
in the area surrounding the sperm head. The pre-coated
 5. Examine halos around the sperm head under
microscope using high magnification ( x40).

Interpretation:
Under microscope halos will be seen around the sperm
heads. The sperm heads showing no halos are
considered negative for acrosomal status and function.

Sperm without halos

Sperm with halos
WARNINGS AND PRECAUTIONS
All human, organic material should be considered
potentially hazardous.
Method
Handle all specimens as if capable of transmitting HIV or
1. Open the diluents ampoule. hepatitis. Always wear protective clothing when handling
2. Add 100 l of well mixed semen sample. Shake a specimens.
little and incubate the mixture for 5 minutes at room
temperature.
3. Apply one small drop of mixture to the pre-coated Cryocell India Pvt Ltd
gelatin film slide and spread smoothly and evenly. A-35, Nirman Vihar, New Delhi 110025 INDIA
4. Allow the fluid to dry partially and then transfer to Tel. No. : (+91 11) 22503735, 22543201
a moistened petri dish (not touching the moistened
tissue) and incubate at 500 C for 30 minutes.