Methods

1. Incubate eggs on their sides for 18-20 h at 38 ±1°C for stages 4 and 5 (3).

2. Wipe or spray eggs with 70% alcohol.

3. Pour cold BSS in pie dish, about 4-cm depth (6-700 mL).

4. Add 1 mL antibiotic/antimycotic solution/100 mL BSS (4).

5. Open eggs with coarse forceps at the blunt end, i. e., air sack.

6. Remove shell to make opening large enough to let yolk out.

7. Pour off thick albumin (into a waste bucket nearby), and save a little of thin albumin in a 90-mm Petri dish or sterile beaker (5).

8. Pour the yolk quickly into BSS (6).

9. Put up to six yolks into the dish, and top up with BSS if they are not completely covered.

10. Remove remaining albumin from all yolks in the dish with coarse forceps or large-mouthed well-flamed pipet.

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