CRYO-NCD KIT

The Nuclear Chromatic Decondensation Test

(NCD test) Principle

The chromatin in spermatozoa is in a highly condensed state prior to fertilization.

The appropriate nuclear chromatin de-condensation and subsequent male

pronucleus formation is essential for fertilization and normal zygote

development.

Highly condensed nuclear chromatin in spermatozoa is because of the formation

of S-S cross links between its histone units. The cleavage of S-S link can be

induced in vitro by Sodium Dodecyl Sulphate and EDTA. Such induced

decondensation is predictor of good fertilizing ability of spermatozoa.

Material included with the Kit

1. NCD Incubation reagent 0.5 ml x 10 ampoules

2. NCD Dry reagent disks 10 nos

3. Fixative and stain reagent 1 ml

4. Parafilm sheets

Material not included with the Kit

Incubator for 50oC

Pipettes

Microscope slides and cover slips

Microscope

Storage and Stability

Store at 2-8 o C till expiry date. Expiry indicated on label

Method:

1. Open one ampoule of NCD Incubation reagent. Add one disk of NCD Dry

reagent to the pipette reagent, cover with piece of parafilm and shake to let

the disk elute.

2. Incubate the vial at 50oC for 5-10 minutes to achieve equilibration at this

temperature.

3. Add 100 microlitre of well mixed semen sample to the incubation mixture

and incubate further for 5 minutes exactly.

4. At the end of incubation time add one drop of fixative and stain reagent to

the incubation mixture.

5. Mix well and take a small drop on slide and place a 22x22 mm cover-slip.

6. Examine under low power and then high power of microscope. The cells

showing swelling of nucleus with uniform chromatin are considered

positive and the cells showing intact nucleus are considered negative NCD.

Results

The result of the test is expressed as the percent swollen cells or NCD reacted

cells to total sperms.

More than 70 % sperms in normal samples show NCD positivity. Less than 70

% NCD positivity is associated with reduced fertility of the sperm.

Warnings and Precautions

- All semen samples should be considered potentially infectious. Handle all

specimens as if capable of transmitting HIV or Hepatitis.

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

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