Sperm Processor SFT - HOS (Hypo-osmotic Swelling Test)

HOS (Hypo-osmotic Swelling Test)

• The intact sperm membrane permits a passage of fluid into the Cytoplasmic space, swells & the pressure generated makes the tail curl.

• All the curled tail sperm are labeled as ‘Live Sperm’.  The damaged / broken sperm membrane, allows fluids to pass across the membrane without any accumulation & hence there is No Cytoplasmic Swelling & the tail remains straight.

1 kit = 100 Tests

HYPO-OSMOTIC SWELLING TEST

HOS (Hypo-osmotic Swelling) assay is based on the principle of osmotic movement of solvents across the membrane. 
Due to the influx of fluid, the sperm tail expands & bulges in a characteristic pattern, which is termed as HOS response.
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Therapeutic Use

• Unlike the dye exclusion vitality assay, the HOS assay do not kill the sperm. Hence, this assay is valued in therapeutic approach to ICSI.

• During ICSI procedure, where only one sperm is used, HOS test allows the technician to pickup viable Sperm. The preferred incubation time herein HOS medium is only five minutes.

Sperm Membrane Integrity & Osmotic Environment

Sperm Membrane Integrity

• It helps in understanding the overall fertilization capacity of sperms.  Compromised sperm membrane integrity has an effect on: Sperm Motility, Sperm Capacitation, Acrosome Reaction & binding of sperm to the Zona Pellucida. 

• Sperm Vitality is measurable through Sperm Membrane Integrity assessment and it reflects the proportion of live spermatozoa.  Sperms exposed to oxidative stress may have DNA damage and are likely to have compromised plasma membrane; thus, are incapable of natural fertilization.

Functional membrane integrity

• Functional membrane integrity is defined as the ability of sperm's outer membrane to maintain equilibrium between the sperm cell contents & surrounding environment. It can be evaluated by response of sperm to osmotic environmental stress.

• Intact sperm membrane permits passage of fluid into the Cytoplasmic space in hypo osmotic environment resulting into sperm tail swelling & curling.  The pattern of curling may vary from sperm to sperm within the same semen sample.

Evaluation of Hypo-osmetic Swelling Test

All the curled tail sperms are considered as Live and Dead Sperms exhibit no cytoplasmic swelling or tail curling (remains straight).

• The sperm tail seems to be particularly sensitive to swelling & curling, as the membrane in this region is loosely attached in comparison to the sperm head area.

• Tail swelling indicates membrane integrity. Curling pattern reflects skeletal integrity and it may vary from sperm to sperm.

• Sperms with intact membrane swell within 5 minutes in HOS medium & all flagellar shapes are stabilized by 30 minutes.

• No tail swelling in majority of the sperms and extreme low motility suggests that the Non-motile sperms are Dead sperm.

• Prior to the HOS test, semen sample should be examined to determine if any sperms have undergone spontaneous swelling or tail coiling. The number of such sperms should be subtracted from the HOS test score in order to obtain accurate results.

Lab Equipments & Accessories

Safety & Environmental Precautions

• Store the kit in appropriate conditions as per the instructions given on the package.

• Do not use the kits beyond expiry date given on the package.

• All the reagents are potentially hazardous and it is strictly advised to follow the MSDS instructions.

• All the patients' samples should be treated as potentially infectious. Laboratory personnel are advised to wear protective gloves, face masks, laboratory coats etc while performing the tests.

• Dispose off all the waste generated, during and after the tests, in an appropriate manner as per the governing compliance laws of the land.

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Test Procedure

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Hypo-osmotic Swelling Test