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 - don't get these anymore - use sodium sulfite instead of soda, shaken water

• club soda
• H2O2

Solutions

• Sodium Bicarbonate 0.8 M
• Sodium sulfite 0.2M
• 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/nc058… 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."
    

From zfin:

Tricaine:

Tricaine (3-amino benzoic acid ethyl ester also called ethyl 3-aminobenzoate) comes in a powdered form from Sigma (Cat.# A-5040). It is also available as Finquel (Part No. C-FINQ-UE) from Argent Chemical Laboratories, Inc. Make tricaine solution for anesthetizing fish by combining the following in a glass bottle with a screw cap:

400 mg tricaine powder
97.9 ml DD water
~2.1 ml 1 M Tris (pH 9).

Adjust pH to ~7. Store this solution in the freezer. (Buy the smallest amount possible because tricaine gets old.)

To use tricaine as an anesthetic combine the following in a 250 ml beaker:

4.2 ml tricaine solution
~100 ml clean tank water.

• 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)

Refractometer

*Reads in Brix. 1degree Brix = % sucrose in solution (m/m)

*ppt saline = ~8 x Brix

• d20/20 (specific gravity) = ~ 0.0064Brix + 1

Glucose Meter

Aimstrip Plus

Fisher Catalog No. 23-111-275

~$18

Should have 1 per group