Page 286 - Proceeding The 2nd International Seminar of Science and Technology : Accelerating Sustainable Innovation Towards Society 5.0

nd
he 2 International Seminar of Science and Technology
“Accelerating Sustainable innovation towards Society 5.0”
ISST 2022 FST UT 2022
Universitas Terbuka

2.6 Semen Evaluation
Semen evaluation was carried out only microscopically for
spermatozoa motility, spermatozoa concentration, spermatozoa
viability and spermatozoa morphology. The assessment is carried out
in the following manner:
a. Spermatozoa Motility
Motility assessment can be done by dripping cement and
physiological NaCl on a glass object with a ratio of 1:4 or 1:5 and
warmed with a heating table. The semen was then observed using
a microscope at a magnification of 400 times by observing the
movement of the spermatozoa. Observations were made in 10
fields of view and graded in %.
b. Spermatozoa Viability
Spermatozoa viability was tested using eosin-nigrosine staining.
The cement was mixed with eosin-nigrosine (1:3) dye. The
mixture of cement and eosin-nigrosine was prepared for smears
and dried on a heating table for 10 seconds. The preparations
were observed under a microscope with a magnification of 40 ×
10. Spermatozoa were observed in 10 fields of view or with a
sperm count of at least 200 cells. The live spermatozoa do not
absorb the color (transparent) and the dead will absorb the red
color on the head. Spermatozoa were counted by dividing the
number of live spermatozoa by the total number of spermatozoa
counted multiplied by 100%, with Equation (1):
Σ live spermatozoa (1)
Spermatozoa viability= × 100%
Σ total spermatozoa
c. Spermatozoa concentration
It was carried out using a counting chamber, carried out by diluting
cement in a formal saline solution with a ratio of 1:200 (5 µl semen:
995 µl formal saline). Observations were made using a
microscope with a magnification of 400 times. Calculations are
carried out on 5 counting chamber boxes, each of which has 16
small boxes. The concentration calculation was done by
multiplying the number of spermatozoa in 5 boxes by 10 .
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256 ISST 2022 – FST Universitas Terbuka, Indonesia
International Seminar of Science and Technology “Accelerating Sustainable
Towards Society 5.0