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Preservation of Wet Anatomical Specimens.

Preservation of Wet Anatomical Specimens. Mr. David Cutting. Technical Officer, Museum of Human Disease, UNSW. davecutting@unsw.edu.au. Old Specimens. Solution may appear discoloured/dirty. -Haemolysis (blood released from tissue). - B ile, mucous or debris in solution.

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Preservation of Wet Anatomical Specimens.

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  1. Preservation of Wet Anatomical Specimens. Mr. David Cutting.Technical Officer, Museum of Human Disease, UNSW. davecutting@unsw.edu.au

  2. Old Specimens. Solution may appear discoloured/dirty. • -Haemolysis (blood released from tissue). • -Bile, mucous or debris in solution. • -Glycerol present in solution may have aged and discoloured.

  3. Old Specimens. What’s in the pot? • Likely to be one of four solutions. Namely Proger’s, Kaiserling or Wentworth’s. • There may be a mix of the three or solutions of varying grade ethanol and formalin dilutions.

  4. Mounting Solutions. Proger’s Solution: to make 11L (10,990mL) Sodium Dithionite (Sodium Hydrosulphite), 90g (0.81%)Di Sodium Hydrogen Orthophosphate, 125g (1.13%)Potassium Di Hydrogen Orthophosphate, 10g (0.09%)Formalin 37-40%, 450mL (4.0%)Pyridine, 90mL (0.81%)Distilled Water, 6.2L (56.4%)Glycerol, 4L (36.3%)

  5. Mounting Solutions. Wentworth’s No.5 Solution: Per 9L (9057mL) Sodium Acetate hydrated, 300g (3.31%)Tri Sodium Orthophosphate, 7.5g (0.08%)Formalin 37-40%, 150mL (1.65%)Distilled Water, 7.5L (82.8%)Glycerol, 1.5L (16.5%)

  6. Mounting Solutions. Anatomy Solution: to make 10LSodium Acetate hydrated, 1.5g (15%)Distilled Water, 5.5L (55%)Glycerol, 3.5L (35%)*sodium dithionite may also be added at 9grm per litre of solution as a colour restorative.

  7. Old Specimens. How can I tell what solution I’m dealing with? • Smell and Viscosity. • The pH is likely to have been basic (7.5) at time of potting but no accuracy after many years. • Specimen records if applicable. • Whatever is in it, it’s likely to be considered flammable.

  8. Old Specimens. How do I remove the old solution? • Open pot, wearing proper PPE in a fumehood or well ventilated area. • Glass/Perspex? • Decant solution into an appropriate container. -Sealable. Non-metallic. Suitable for corrosives.

  9. Old Specimens. Once the solution is removed. • Handle the pots very carefully as not to dislodge and damage the specimen. • Rinse with water. • Clean out inside with a detergent like ‘Pyroneg’ or ‘Sonidet’. • Keep loose specimens in preservative.

  10. Old Specimens. How do I dispose of the old solution? • Adhere to your facility’s waste disposal guidelines. • Label the waste appropriately.-Ethanol/formalin waste. • Check with your OHS/Waste Management departments. • Don’t pour it down the sink!

  11. Replacing fluid. What preservative solution do I use? • 1. Proger’s.2. ‘Anatomy’3. Parraffin Oil. 4. Wentworth’s No.5.

  12. ‘Proger’s’ Mounting Solution. • Not suitable for fatty specimens. Will break down fat. • Many components, lengthy to make. • Most toxic solution. Pyridine/Sod. Dithionite/Formalin. + Excellent as a colour reclamation solution for old, faded specimens. + Anti-fungal qualities. + Will maintain clarity for longest.

  13. ‘Anatomy Formula’ Mounting Solution. • Impurities in Sodium Acetate can affect refractive index and appear cloudy. • No real anti-fungal properties. • No colour restoration qualities.* + *Sod. Dithionite may be added (9grm per L) + Easy to make, few components. + Very low toxicity.

  14. Paraffin Oil. (light grade) • Poor colour reclamation. • Cost. • Requires specimen and pot to be dry (immiscible with H20) which can damage tissue is over dried. • Not suitable for thin-walled specimens. E.g. Intestinal tract. + Will not encourage leeching of aqueous pigments from tissue. + Great clarity of solution. + Inert, non-toxic. + Does not support mould growth. + No preparation required, can be used straight from bottle.

  15. ‘Wentworth’s No.5’ Mounting Solution. • More suited to new specimens, following fixation in Wentworth’s No1. Solution. • Poor colour reclamation*. • Can tend to create a concavity in perspex pots. + Versatility. Can be used with all tissues. + Low toxicity.

  16. Repaired Specimens. Now that your specimens are cleaned and looking refreshed... • Keep them out of direct sunlight. This will bleach the tissue. • If in perspex, handle carefully and do not squeeze the pot. • Keep records of which solution they are kept in. • Check regularly for any leaks, changes in the fluid or visible damage to the pot.

  17. To order new pots, see http://www.plaztekscientific.com.au/ or any local perspex wares manufacturer. There was a very good company in Canberra called Austral Scientific but they seem to have disappeared. • For chemicals and materials, seehttp://www.livingstone.com.au/http://www.vitalmed.com.au/http://ajaxfinechem.com/http://www.sigmaaldrich.com/australia.html • Feel free to email me with any potential problems you may encounter. My address is in the first slide of the presentation. Good luck.

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