Comparative Physiology @ Guelph
Slime Collection Protocol - Myxine glutinosa
Slime Collection Protocol – Myxine glutinosa
Materials:
- 3.5 L fresh, artificial seawater, 10 degrees
- clove oil anaesthetic stock (1 part clove oil: 9 parts anhydrous ethanol)
- bucket of ice
- squirt bottle of dH2O
- 100 mL beaker of dH2O
- narrow spatula for scooping slime
- vial(s) of stabilization buffer on ice
- paper towels
- Kimwipes
- Large dissection tray filled with ice
- Small dissection tray lined with clean J-cloths soaked in s.w., wrung out
- Electrical stimulator and stimulator wand
- Plastic bag with ice for regulating water temperature
Procedure:
- Add 1.75 mL of clove oil stock solution (1:9 co:ethanol) and mix
- Transfer to white bucket for trip to Aqualab
- Bring a spare bucket for recovery water
- Rinse dipnet thoroughly (*very important*), collect hagfish as gently as possible
- Transfer hagfish into anaesthesia bucket, close lid, note time
- Collect a couple of litres of water from hagfish tank into recovery bucket
- Bring hagfish and recovery water back to lab
- Set up dissection trays
Make sure stimulator settings are correct: Frequency: 60 Hz, Delay: n/a, Duration: 0.8 ms, Volts: 18 V, Mode: repeat, Output: bi *NOT DC or mono*
- Hagfish should be knocked out after 10 minutes or so in bucket
- Once hagfish stops moving, wait another couple of min. to ensure complete anaesthesia
- Place hagfish in chilled tray, ventral side up
- Rinse area where slime will be collected
- Blot dry with Kimwipes
- Press firmly with stimulator wand at ventral midline between opposite slime glands
- Remove wand before exudate contacts it
- Scoop up exudate with spatula, transfer to stabilization buffer, twirl to disperse
Wipe off spatula, rinse in dH2O, wipe again to dry *This is important to ensure that stabiliation buffer does not contact the hagfish’s skin*
- When finished with slime collection, return hagfish to recovery bucket, massage skin with gloved hand to remove any adherent slime. Return to Aqualab ASAP.