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(Source:
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D.
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Frost
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and
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D.
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Sepich
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)
Zebrafish embryonic cells attach well to glass coated with laminin or to plastic. They do not attach well to bare glass or glass coated with fibronectin. They attach somewhat to 0.2 mg/ml gelatin coated glass.
- Sterilize glass coverslips (22 mm x 22 mm) by dipping in ethanol and flaming.
- Place a drop of 10 μg/ml laminin (Boehringer Mannheim) in sterile PBS and leave for 45 minutes.
- Rinse thoroughly with sterile PBS or dH2O.
- Place a drop of cell suspension (cells from 1-4 embryos) onto a 0.5-1.0
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- cm
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- spot
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- in
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- the
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- center
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- of
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- the
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- coated
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- glass
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- coverslip
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- in
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- a
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- Petri
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- dish
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- and
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- allow
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- the
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- cells
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- to
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- attach
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- for
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- several
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- hours
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- or
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- overnight.
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- Flood the dishes with growth medium.