Fish labs

Lab 1: Learn to use the oxygen equipment

Per table:

  • 4 50 mL beakers
  • 250 mL flask
  • 100 mL beaker
  • parafilm
  • kimwipes
  • paper towels
  • dissecting tray
  • RO water in squirt bottle

Shopping

  • club soda
  • H2O2

Solutions

  • Sodium Bicarbonate 0.8 M
  • Sodium sulfite (don't have as of 2017)
  • Sodium carbonate 0.8 M

Lab 2

Try one of 12 protocols approved by IACUC

Lab 3

  • anesthetic (see paper MS222 MESAB, lidocaine) See paper here: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3966278/
    • lidocaine http://www.spexcertiprep.com/search.aspx?search=lidocaine 1mg/mL, FISHER NC0589195. "Lidocaine hydrochloride (Lidocaine Hydrochloride 10% for Injection USP, Hospira, Lake Forest, IL) was instilled beneath the surface of 1 L of system water in a 2-L tank by using a 25-gauge needle and stirred into solution to achieve final concentrations of 300, 325, and 350 mg/L at pH 6.9 to 7.4. Fish were added directly into the tank with the prepared lidocaine hydrochloride solution." [We had to buy 1 mg/mL in methanol]
    • MS222 (tricaine-S, MESAB0 https://punchout.fishersci.com/shop/products/tricaine-s-ms222-10g/nc0589195 FISHER 2347) "MS222 (Finquel Tricaine Methanesulfonate, Argent Chemical Laboratories, Redmond, WA) prepared under a chemical fume hood was mixed 1:2 with 95% sodium bicarbonate to make a stock solution of 10 mg/mL with a resulting pH of 7.3 in system water. The stock solution was added to the anesthetic tank containing system water to achieve a final concentration of 150 mg/L, to which the fish were added."
  • warm (37C water bath)
  • razor blades
  • glass plate to cut on
  • fixative (paraformaldehyde in buffer - get from Rolf's lab)
  • stirplate & stirbar
  • thermometers
  • ice bucket
  • noise (Rolf brings speaker)
  • bright light
  • sodium sulfite
  • capillary tubes
  • glucose strips
  • salinity (5M NaCl)
AttachmentSize
3.zebrafishmetabolismlabmanual2017.v5r.pdf1.19 MB