Manufacturers of:

IUI Media
ART Equipments
ART Consumables
Frozen Donor Semen Sample

Interpretation
Spermatozoa that are white (unstained) are counted as live and those
showing any degree of pink or red are dead.

Limitations of the method

Spermatozoa stained with this kit can not be used for any further
procedures. To differentiate live spermatozoa from dead spermatozoa for
use in ICSI (low viability samples), the HOS (hypo-osmotic swelling) test
should be used.

Warnings and precautions

Contains:

Cryocell India Pvt. Ltd.

Reg. Office:
A-35, Nirman Vihar, Delhi-110092
Works:
1-11-153, Sindhi Bazar, Jalna - 431203, (M.S.) India
info@cryocellindia.com, cryocellindia@hotmail.com
www.cryocellindia.com

Cryo-Vitality EN & E: 1 ml

CRYO-Vitality E & EN
Sperm Vitality Staining Kits

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.

Store at 2C - 8C
For Research Use Only

The sperm vitality is reflected in the proportion of spermatozoa that are

alive. It is measured by assessing the ability of sperm plasma
membrane to exclude extra-cellular substances like dyes. Sperm vitality
should be determined in semen samples with less than fifty percent
motile spermatozoa. Vitality assessment also provides check on the
accuracy of motility assessments; as the percentage of live
spermatozoa should be slightly exceed the total percentage of motile
spermatozoa.
Eosin-Nigrosin staining is used for assessing vitality. The technique is
based on the principle of dead cells will take up the eosin, and as a
result stain pink. The Nigrosin provides a dark background, which
makes it easier to assess the slides. The assessment can be carried out
at any time and slides also can be preserved for future assessment and
record.
Plain eosin staining also can be used to assess vitality in wet smears.
This provides quick assessment at the same time of count and motility
assessment.

METHOD
Cryo-Vitality EN

Material included in the kit

Cryo-Vitality EN
Cryo-Vitality E

1 ml
1 ml

Storage
Store reagents at 2-8C

Equipment required
Light microscope (1000X magnification)
Microscope slides pre-cleaned
Cover slips
Pipettes
Test tubes

1. Mix equal volumes of well-liquefied semen and Cryo-Vitality EN (small

drops) on a pre-cleaned microscopic slide. Allow about 30 seconds for
staining.
2 Make smear on the slide using other slide as spreader. Smear should be
thin enough for allowing proper visualization. Thick smears are useless
for evaluation.
3 Allow air-drying and preferably mounting with DPX or equivalent
mountant as soon as possible. These slides can be stored at ambient
temperature indefinitely.
4 At least 100 or preferably 200 spermatozoa are examined at a
magnification of 1000x or 1250x under oil immersion using high quality
non-phase-contrast objective and correctly adjusted bright-field optics.

Cryo-Vitality E
1 Mix equal volumes of well-liquefied semen and Cryo-Vitality E (small
drops) on a pre-cleaned microscopic slide. Cover with a 22x22 mm
cover-slip If there is excess mixture of semen and stain seen blot lightly
with tissue paper. Allow 5 minutes for settling and staining of the
sperms.
2 Examine under high power using high quality non-phase-contrast
objective and correctly adjusted bright-field optics.