Acrosome function test GELATIN FILM LYSIS TEST KIT

The sperm acrosome is located on the top of the sperm

head. During acrosome reaction, the outer acrosomal

membrane and the plasma membrane fuse and vesisculate

to discharge the acrosomal contents. Multiple enzymes are

present in the sperm acrosome including several acid

hydrolases commonly found in lysosomes. Acrosomal

enzymes help the sperm to penetrate the cumulus mass and

the zona pellucida

A sperm with defective acrosome may not have the

sufficient stock of these proteolytic enzymes and would

eventually fail to penetrate the zona, a prerequisite for

successful fertilization.

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

gelatin film on the glass slide is lysed coming in contact

with proteolytic enzymes leaching out of acrosome and

allow light to pass through creating a halo around the

sperm head which can be examined using a microscope.

Material included with the Kit

1. Incubation reagent.

2. Pre-coated Gelatin Film Slides.

Material not included with the Kit

1. Plastic vials

2. Pipettes

3. Incubator at 50oC.

4. Petri dish with moist filter paper

Storage and Stability : Store at 2-8

O C till expiry indicated on label.

Sperm without halos

Sperm with halos

Method

1. Pipette 0.5 ml of Incubation reagent in a plastic vial.

2. Add 100 µl of well mixed semen sample. Shake a

little and incubate the mixture for 5 minutes at room

temperature.

3. Apply one small drop of mixture to the pre-coated

gelatin film slide and spread smoothly and evenly.

4. Allow the fluid to dry partially and then transfer to

a moistened petri dish (not touching the moistened

tissue) and incubate at 500 C for 30 minutes.

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.

WARNINGS AND PRECAUTIONS

All human, organic material should be considered

potentially hazardous.

Handle all specimens as if capable of transmitting HIV or

hepatitis. Always wear protective clothing when handling

specimens.

Cryocell India Pvt Ltd A-35, Nirman Vihar, New Delhi 110025 INDIA

Tel. No. : (+91 11) 22503735, 22543201