Page 285 - Proceeding The 2nd International Seminar of Science and Technology : Accelerating Sustainable Innovation Towards Society 5.0

nd
The 2 International Seminar of Science and Technology
“Accelerating Sustainable innovation towards Society 5.0”
ISST 2022 FST UT 2022
Universitas Terbuka
2.2 Preparation of Diluent
All chemicals used were Merck products (Germany), and antibiotics
(Meiji Seika Pharma, Japan). Tris basic diluent composition (buffer) is
presented in Table. 1.

Table 1. Tris Buffer Composition
Composition Amount
Tris (hydroxymethyl) 3.03
aminomethane (g)
Citric acid (g) 1.78
Fructose (g) 1.25
Aquabidest (ml) 100
The buffer was then added with 20% egg yolk from the total volume of
the diluent, 6% glycerol and 1000 IU/ml antibiotics and 1 mg/ml
streptomycin. The diluent is frozen and thawed at the time of use.
2.3 Testicular/Epididymal Collection
Testicular/epididymal collections were carried out at RPH Bogor,
Yogyakarta and Bali. The testes and epididymis were taken from
males that was just slaughtered 15 minutes before.
2.4 Organ Preparation
The testes were prepared up to the vas deferens, both ends of the
sections were ligated to prevent the release of spermatozoa. The
connective tissue around the organ is removed carefully to avoid
bleeding. Organs were taken and immersed in physiological NaCl
solution (0.9% NaCl). The testes and epididymis were measured for
parts.
2.5 Spermatozoa Collection
Spermatozoa were collected from dexter and sinister. The head, body
and cauda were incised alternately using a scalpel. Each part of the
epididymis is flushed with a cement diluent solution. The white liquid
that comes out of each part of the epididymis is collected in a petri
dish.

ISST 2022 – FST Universitas Terbuka, Indonesia 255
International Seminar of Science and Technology “Accelerating Sustainable
Towards Society 5.0