ON TESTIS AND EPlDlDYMlS OF RATS - Pondicherry University ...

1.1 Preparation of dosing solution
APPENDIX 1
PREPARATION OF REAGENTS
1.1.1 2.3.7.8-Tetrachlorodibem-p-dioxin (TCDD)
100 pg of TCDD was dissolved in acetone and olive oil (1:19) and used as
stock solution. The stock solution was serially diluted and used for treatments.
1.1.2 Vitamin E
200 mg of vitamin E was made up to 10 mL with olive oil and stored in amber
coloured bottle at 4OC. This solution contained 20 mg vitamin E/ mL.
1.2 Epididymal sperm counts
1.2.1 Diluents
The diluents for sperm counts were prepared by the addition of 50 g sodium
bicarbonate, 10 mL 35% formalin and 0.25 g trypan blue in a volumetric flask
and made up to a final volume of 1 L with distilled water.
1.3 Serum FSH
1.3.1 Anti-FSH-Coated microtitration strips
One strip holder contained 96 microtitration wells coated with anti-FSH
antibody. Stored at 2-8'C in the resealable pouch with a desiccant to protect
from moisture.
1.3.2 FSH standards
One vial, 2 mL, labeled A contained 0 mlU1 mL and five vials. 1 mL each,
labeled B-F, contained concentrations of approximately 1.5, 4.5, 15, 45 and
150 mIU/ mL FSH (from human pituitary gland) in porcine serum with a nonmercury
preservative.
1.3.3 FSH controls
Two vials, I rnL each, Levels I and 11. contained low and high concentrations
of FSH (from human pituitary gland) in porcine serum with a non-mercury
preservative.