NBP

NBP kdorfman Mon, 01/16/2017 - 14:40

Bio 397NP Neurobiology and Physiology Lab

Color vision

Color vision kdorfman Thu, 04/13/2017 - 14:24

Fish labs

Fish labs kdorfman 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

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

Mussel projects

Mussel projects kdorfman Wed, 02/08/2017 - 22:21
  • Make 2 L Instant Ocean, aliquot
  • Change pH with NaOH, HCl, see how long it lasts
  • hot block or digital hot plate
  • order epinephrine (bought 1 g from Acros, should try solution: Fisher S25126 L-Adrenaline Solution, 0.01% = 0.01g/100 mL = 0.1 mg/mL, 100 mL $6.37) solubility = 0.18 mg/mL
  • Make 10X acetylcholine, serotonin
  • Check ATP supply
  • find thermometers
  • vasopressin
  • nicotine (10^-3, 10^-4, 10^-5, 10^-6 in instant ocean)
  • Lactic acid

F2017 Mussels

F2017 Mussels kdorfman Mon, 09/18/2017 - 18:53

Projects

  • 2L Instant Ocean

  • CaCl

  • Adrenaline Cl 1:10,000, 100 mL (0.1mg/mL)

    • Fisher S25126
    • L-Epinephrine CAS# 51-43-4
    • 183.207 g/mol
    • 180 mg/L (at 20 °C). Very slightly soluble in alcohol. Readily soluble in aqueous solutions of mineral acids, NaOH, and KOH. Insoluble in aqueous solutions of ammonia and of the alkali carbonates. Insoluble in chloroform, ether, acetone, oils.
  • Creatine Phosphate

    • Fisher AC 337340010 1 g $55.95
    • MW 321.14
    • H2O: 0.1 g/mL, clear, colorless
    • 32 mg/1 mL =~100mM
  • Nicotine

    • Fisher AC181420050
    • 99%
  • Serotonin

  • acetylcholine