Comparative Physiology @ Guelph
Hagfish Slime Separation
Prepare 500mL of stabilization solution and chill to 4 degrees Celcius before beginning the slime separation procedure.
Collect desired amount of fresh exudate into chilled stabilization solution.
- Millipore filtration system (upper portion, lower stem without white insert, metal insert for stem, and 53um mesh cut to size)
- 2 x Erlenmeyer flasks
- Falcon tubes for sample storage.
- IMPORTANT: All equipment that will come into contact with thread cells and mucins must be thoroughly washed and rinsed with ultrapure water. The equipment must also be RINSED WITH STABILIZATION SOLUTION to prevent rupture of mucins.
- Assemble the filtrate system and fit onto one of the flasks
- Re-suspend the slime sample that has been collected into stabilization solution. Pour this through the filtration system.
- The filtration rate can be controlled by adjusting the seal between the rubber stopper and the flask. It is VERY IMPORTRANT that the thread cells are not exposed to the air.
- Continue to rinse thread cells with chilled stabilization solution (3-4X).
- If the mucin vesicles are going to be used it is important that they remain fairly consentrated to do this the filtration system can be switched from one flask to the other and the filtrate containing mucins poured into individual falcon tubes.
- When satisfied with the separation the filtration system (not the flask) can be inverted and rinsed with stabilization solution to get the thread cells into a falcon tube.
- After cleaning up the equipment and rinsing again with stabilization solution the samples can be re-filtered to remove any clumps of thread cells or mucin vesicles respectively. The procedure is the same however a 300um mesh can be used on the thread cell sample to remove any clumped thread cells.
- Repeat as necessary.
Written by Tim Winegard posted 11/27/08