Effects of isochaborat testosterone on the kidney rats

Materials and Methods
2.5.Method used in perfusion: (Humason1979)
The equipment used in perfusion comprises: two 2000 ml glass flasks
(for the saline and the fixative respectively), each equipped with an outlet at
bottom, elevated 1.2 meter above the animals; two sets of plastic tubing with a
flow regulator; a plastic” Y” connector tubing and the connection were kept free
of air bubbles.
After anaesthesia, the animals were attached to a perfusion tray by
means of masking tape. The thorax was opened to expose the heart. An incision
in the left ventricle was made and the cannula was inserted in the ascending
aorta through the left ventricle. The cannula was clamped inside the aorta with a
haemostatic clamp.The right atrium was opened and the saline was allowed to
flow at a maximum rate for 30 seconds to 2 minutes (180mm Hg)
The perfusion with the fixative was started maximum rate and the flow
of the saline was decreased to zero. After 1~2 minutes the perfusion rate was
slowed down and maintained at a pressure of 100 mm Hg for 30 minutes.
2.6. Post-fixation, dehydration and embedding:
The kidney was cut into 2-3 mm segments, immersed in fresh fixative at
4 ºC for further 24 hours. The segments were then washed in 0.1 M cacodylate
buffer two times 10 minutes each and then transferred into a secondary fixative
consisting of 1% osmium tetraoxide in 0.1 M cacodylate buffer for afuther 2 h
(on ice). After this, they were dehydrated through graded alcohol series, cleared
in propylene oxide, and finally embedded in Epon 812.
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