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Table 1
Hematocrit levels in light-damaged rats
HCT (%)
LD Ctrl
47.2 ± 1.3 (24)
EPO
S100E
LD IM
71.5 ± 5.7 ( 10)*
52.3 ± 5.2 (10)*
LD SR 57.8 ± 7.9 (10)* 49.8 ± 3.7 (10)
Ctrl uninjected rats, HCT hematocrit, LD light-damaged rats, IM intramuscular injec-
tion, SR subretinal injection. The measurements were performed at P65. The values are
means ± SD, the number of eyes analyzed is shown in brackects ( 46 ). The asterisk
indicates hematocrits significantly increased (t-test p<0.05) compared with controls.
Reproduced from Colella P. 2011 ( 46 ) by permission of Oxford University Press
1. Following blood and ACF collection (see Subheadings 3.6.2
and 3.6.3 ), sacrifice the rat using a lethal dose of Avertin or by
cervical dislocation.
2. Mark the temporal side of the eye searing the sclera with a
cauterizer (see Note 38).
3. Enucleate the eye making several incisions around the globe
with scissors. Be gentle to avoid damaging the eye.
4. Put the eye in a test 2 mL tube containing 4% PFA for 12 h at
4°C.
5. Wash the eye for 5 min twice in 1× PBS.
6. Dehydrate the eye in 30% sucrose until the eye drops to the
bottom of the 2 mL tube (usually about 4-8 h for a rat eye).
7. Take the eye with tweezers and lean it on a paper to remove
dripping sucrose.
8. Fill in a plastic mold with tissue freezing medium (O.C.T.).
9. Lay the eye on one side at the bottom of the mold. Lay the eye
so that neither the anterior nor posterior part of the eye faces
the bottom of the mold (see Note 39).
10. Mark the side of the mold (left or right) that corresponds to
the temporal side of the eye (see Note 40).
11. Allow the O.C.T. to solidify putting the mold in a bath with
dry ice and denatured ethanol.
12. Store the mold at −80°C.
3.6.4 Eye Harvesting
and Embedding
 
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