Chick agar culture

In vitroculture of early chick embryos

1. Use sterile technique. Prewarm Howard's Ringers in petri dish and
agar/albumin culture dish to 37ºC.

2. Crack 2-day egg into large sterile petri dish. Alternately, open blunt end
of egg and remove shell membrane. Use disposable pipet to remove some
of the albumin, so that top half of yolk is uncovered. Place filter paper
ring around blastodisk, and cut around it. Transfer filter paper ring with
embryo to warm Howard's Ringers. Stage embryo. Gently wash away
adherent yolk.

3. Transfer to agar/albumin dish filter paper-side down (ventral side of
embryo is up).

4. Incubate in 37º incubator overnight. Check for evidence of development
the next day, and thereafter.

Albumin-agar plates

Ref. Biroc, S., Developmental Biology, a Laboratory Course with Readings,
Macmillan Publishing Company, New York, 1986 , p.48

1. Autoclave 200 ml beaker with stir bar. Wipe eggs with 70% ethanol. Collect 100 ml albumin by separating egg whites from approximately 5 eggs into sterile beaker (discard yolks). Stir at room temperature to break up clumps. Warm to 45ºC in water bath.

2. Autoclave 1.3 g agar, 1.3 g glucose and 33 ml Howards' Ringers solution in 100 ml flask for 15 minutes. Cool to 45ºC in water bath.

3. Combine agar and egg whites with stirring. Transfer between flask and beaker to aid combination. Turn water bath up to 48ºC to keep agar from solidifying while pouring plates.

4. Cut tip off of transfer pipet. Transfer approximately 3 ml to 35 mm petri dishes. Avoid transferring bubbles.

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		**In vitroculture of early chick embryos** 1. Use sterile technique. Prewarm Howard's Ringers in petri dish and agar/albumin culture dish to 37ºC. 2. Crack 2-day egg into large sterile petri dish. Alternately, open blunt end of egg and remove shell membrane. Use disposable pipet to remove some of the albumin, so that top half of yolk is uncovered. Place filter paper ring around blastodisk, and cut around it. Transfer filter paper ring with embryo to warm Howard's Ringers. Stage embryo. Gently wash away adherent yolk. 3. Transfer to agar/albumin dish filter paper-side down (ventral side of embryo is up). 4. Incubate in 37º incubator overnight. Check for evidence of development the next day, and thereafter.

		Albumin-agar plates Ref. Biroc, S., Developmental Biology, a Laboratory Course with Readings, Macmillan Publishing Company, New York, 1986 , p.48 1. Autoclave 200 ml beaker with stir bar. Wipe eggs with 70% ethanol. Collect 100 ml albumin by separating egg whites from approximately 5 eggs into sterile beaker (discard yolks). Stir at room temperature to break up clumps. Warm to 45ºC in water bath. 2. Autoclave 1.3 g agar, 1.3 g glucose and 33 ml Howards' Ringers solution in 100 ml flask for 15 minutes. Cool to 45ºC in water bath. 3. Combine agar and egg whites with stirring. Transfer between flask and beaker to aid combination. Turn water bath up to 48ºC to keep agar from solidifying while pouring plates. 4. Cut tip off of transfer pipet. Transfer approximately 3 ml to 35 mm petri dishes. Avoid transferring bubbles.