Egg Preperation for Eye Experiments-snippet
From CISSTwiki
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Keywords
Egg, Experiment, Chicken, How to, guide
Description
Preparation of Eggs for Eye Experiments
- For vein cannulation should be 12-14 days in age for best results
- Hatches 19-22 Days
- Euthanize by placing in the freezer.
- The specimen should last about 2 hours with proper �watering�
Need:
- Egg holder
- Buffer solution
- Pipette
- Cup for shell pieces
- Tweezers used for electronics work
- Paper towels
1. First, break at air pocket at top of egg (put egg under light to find)
2. Poke a hole with the tweezers in the top of the air pocket and slowly peel shell off (in small pieces); get as close to air-egg margin as possible without passing/ripping. If you go below, the membrane rips and it will bleed. These last few millimeters is the toughest part.
3. Clear pieces of shell that might have fallen inside, blow them away, or pick up gently.
4. PBS - Solution (14190136 from invitrogen.com - Dulbecco's Phosphate-Buffered Saline solution , chem dept has some) use dropper to moisten the membranes. This makes the membranes easier to separate. Try dropping on side where you may have punctured the membrane a bit. This would make the solution go between the layers and separate them more effectively. Keep the membrane moist throughout � dry=bad. Wait a few minutes after initial application.
5. Don�t forget to add solution to prevent the membrane from drying out.
6. Use the tweezers to try to grab the top membrane � it is extremely thin, so it might take a few tries to grab it (be super careful!). Try to remove large pieces if possible when peeling. Remove all of the top (air pocket) membrane.
Notes:
- Can see the blood vessels clearly � even those beneath the top layer, will bleed if punctured.
- Membrane separating chick and air � 2 thin layers (we are removing the top layer)
- Older specimen, ie 15 days - membranes stick together more it might bleed as you peel the top layer off
- Incubator should be at 37C, and each specimen should be labeled with its age on upon delivery.
- Injector flow rate should be 0.001ml/min or slower?
- Bubbles in the oil should be avoided (may block flow). The oil will automatically flow when vessel is cannulated.
Attachment
egg preparation with images.doc
If you need other animals, ie pigs see:
http://www.cisst.org/wiki/Specimen_for_Experiments_Source-snippet
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