[
Worm Breeder's Gazette,
1977]
In the meeting, some persons were interested by my stained gonads slides. This technical note describes how I proceed to get them. DISSECTED WORMS Fixative = sublimate acetic : 95 ml sublimate (dissolve 10 g of HgCl2 in 100 ml boiling water-let decant one night) and 5 ml acetic acid. When dessication of dissected worms is completed, deposit a fixative drop on the frottis, then immerse the slide in sublimate acetic for 10 mn. Rinse in water and let one (or more) night in iodine-iodized alcohol (100 ml 70% ethanol, 2g I2, 4g KI). Rinse in a 0.25 % sodium hyposulfite solution (5 mn), then in running water (10 mn). Immerse in HCl for 12 mn in a 56 C water-bath, stop hydrolysis by a room temperature water rinse, immerse in Schiff reagent for 2 hours and rinse in running water (15 mn). Stain finally by a fast passing over in : - 1% light green solution, rinse and dehydrate in 1 x 70, 1 x 95, 2 x 100 alcohols, then 2 x Toluene and mount. - picro indigo-carmine (0.25 g indigo-carmine in 100 ml of picric acid aqueous saturated solution). Rinse directly in 100 ethanol ( twice). I prefer use P.I.C. to light green, because the green staining disappears from cytoplasm in 2 to 3 months ENTIRE WORMS (for sections) Fixative = Zenker fluid - stock solution = HgCl2 : 5 g; K2Cr2O7 = 2. 5 g; Na2SO4 = 1g; distilled water 100 ml - Before use, add 5 ml acetic acid. Background staining = Heidenhain's blue - Dissolve in 100 ml hot