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Part 1. Set up

• Dissecting microscopes
• Compounding microscopes
• 7-10 day old wild type zebrafish (1/student minimum)

For 2 stations, 25 students each:

• 30 mm diameter disposable Petri dishes (2/student minimum)
• Tricaine solution (0.4% 3-aminobenzoic acid ethyl ester, pH7)
• Glass slides (1/student minimum)
• Glass coverslips, 22 mm x 22 mm, #1 (7/student minimum)
• Probes for manipulating fish (1/student minimum) [see procedure for making probes]
• Microcentrifuge tubes, disposable, 1.7 ml (1/student minimum)
• Pipettes, glass controlled drop disposable, 5.5" long (15+/student minimum)
• Rubber bulbs (1/pipette)
• Beakers, 250 ml (10)
• Bottles, 100 ml (10)

Solutions:

(staining protocol adapted from Ruth BreMiller)

For Each Station:

• One 50 ml 0.5% TCA in 100 ml bottle with a pipette and bulb and a 250 beaker labeled "TCA waste"
• Two 25 ml Acid-Alcohol in 100 ml bottle with a pipette and bulb and a 250 beaker labeled "Acid-Alcohol waste"
• One 50 ml Alcian in 100 ml bottle with a pipette and bulb and a 250 beaker labeled "Alcian Blue waste"
• One 50 ml glycerol-KOH in 100 ml bottle with a pipette and bulb.

Procedure for Making Probes

These are made by using 6" lengths of 1.2 mm OD glass electrode capillaries pulled apart over a Bunsen burner flame to make 2, approximately 3" long, tapered tip probes. The opposite ends are simply fire polished to make a smooth end.

Go to Part 2
Go to Laboratory Protocol page