Standard Embryo Collections

1. Polystyrene Petri dishes (60 x 15 mm [Falcon, no. 351007], 150 x 15 mm [Falcon, no. 351058]). These need not be sterile, but can be washed and reused.

3. Apple juice (any standard grocery store brand will do).

4. Table sugar (standard grocery table sugar).

5. Nipagin (p-hydroxybenzoic acid methyl ester, Sigma-Aldrich, no. H6654) 15% in ethanol.

6. Yeast paste (standard grocery baker's yeast dissolved in warm water to form a paste; store at 4°C).

7. Population cage: acrylic wide-mouthed (to accommodate 150 x 15-mm plates) cage of approximate dimensions of 16 x 16 x 16 in.; can be custom-ordered from Owl Scientific Co. or constructed by most in-house shop facilities.

8. Collection bottles: 100 mL Tricornered beakers (Fisher no. 02-593-50B) will fit 60 x 15-mm collection plates. For collections on consecutive days, the beakers will become too wet, so either change flies into fresh beakers daily or cut off the bottom of the beaker and affix a nylon filter to permit airflow.

9. Nylon Spectra/Mesh filters, 70 |im (Fisher, no. 08-670-199).

10. Collection basket: two screen cups (Sigma-Aldrich, no. S1145) and 40 mesh screens (Sigma, no. S0770). Insert a Spectra/Mesh filter above the mesh on one of the baskets.

11. Paintbrush.

12. Distilled water (dH2O) Squirt bottle.

13. Clorox (dilute 1 : 1 with dH2O just before use).

14. Plastic weigh boat (Fisher, no. 02-202B).

15. Glass test tube with screw cap (Fisher, no. 14-930AA).

16. Heptane. (Do not breathe vapor or get on skin; highly flammable.)

17. 9-in. Pasteur pipets.

18. Phosphate-buffered saline (PBS): 0.9% NaCl, 14 mMNa2HPO4, 6 mM NaH2PO4, pH 7.3.

19. Fixative: 4% Paraformaldehyde in PBS. Dissolve 4 g paraformaldehyde in 100 mL PBS by adding 10 N NaOH dropwise with stirring until dissolved. Then, add 1 N HCl to bring to pH 7.0. Store at 4°C for up to 1 mo. (Highly toxic; wear gloves and avoid direct contact or inhalation of powder.)

20. Bouin's Fluid fixative: 0.66% picric acid, 9.5% formalin, 4.7% acetic acid. (Highly toxic; wear gloves and avoid direct contact or inhalation.)

21. Methanol containing 5 mM EGTA (pH 8.0). (Avoid contact, inhalation, or ingestion of methanol as it is poisonous and may cause blindness.)

22. PBST: PBS containing 0.4% Triton X-100. Store at 4°C.

23. Normal goat serum (NGS) (Sigma-Aldrich, no. G6767). Store at -20°C.

24. 5% Sodium azide stock solution (dilute 1:100 for working solution). (Highlypoisonous; avoid contact, inhalation, or ingestion.)

25. Blocking buffer: PBST containing 1% NGS and 0.05% sodium azide. Prepare fresh.

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