Fish labs

Submitted by kdorfman on Tue, 02/21/2017 - 20:34

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


  • 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:
    • 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… 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 (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)


*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


Should have 1 per group