Overview of Otolith Thin Slice Method

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Overview of Otolith Thin Slice Method. SDAFS Catfish Technical Committee 2014 Catfish Aging Workshop. Procedures. Remove and dry Set in epoxy resin Thin slice using a slow speed precision saw Mount thin slice to a slide using Cytoseal ™ - PowerPoint PPT Presentation

Transcript of Overview of Otolith Thin Slice Method

Overview of Otolith Thin Slice Method

Overview of Otolith Thin Slice MethodSDAFS Catfish Technical Committee2014 Catfish Aging Workshop

ProceduresRemove and dry

Set in epoxy resin

Thin slice using a slow speed precision saw

Mount thin slice to a slide using Cytoseal

View using a fiber optic light and dissecting scope

List of Items Used For Thin Slice MethodSlow speed precision saw (Isomet 1000) and cutting fluid (Isocut Plus Cutting Fluid)

Epoxy resin and hardener (West Systems 105 epoxy resin and 207 hardener)

Ice cube trays

Disposable plastic cup and spoon

Microscope slides

Cytoseal 60

Forceps

Warm water

Paper towels

2. Set otolith in epoxy resin

-Mix resin and hardener in a disposable cup-Pour base layer of epoxy and let harden (~4 hours)-Set otoliths in the tray-Cover with a second layer of epoxy resin and let harden (~typically overnight)

2. Set otolith in epoxy resin

Set otoliths in resin with posterior edge near the edge of the epoxy resinBe Consistent! Set all otoliths with the same orientation to be able to make the same, consistent, cut on each otolith.

3. Thin slice w/slow speed precision saw

3. Thin slice w/slow speed precision saw

Align the saw bladeslightly to the right of the core of the otolith

Zero the micrometer, setspeed to 200 , lift chuck counter balance and pushthe blade button 3. Thin slice w/slow speed precision saw

Cut off the posterior end of the otolithRaise the counter balance, spin the micrometer wheel until .025 appears on the digital readout3. Thin slice w/slow speed precision saw

3. Thin slice w/slow speed precision saw

Rinse thinslice in warm waterand dry

Repeat process to take a second cut

4. Mount thin slice to a slide using Cytoseal

Apply a drop of Cytoseal to theslide, place thin slice on slide and cover with CytosealRemove air bubbles Allow to dry ~ 1 hour to viewboth sides of thin slice5. View

5. View

5. View

5. View

Same technique can be used for many speciesFlathead catfishStriped bassLargemouth bassBlack crappieGrass Carp??

Questions