Donor Tissue Kindly Provided by Dr. Gloria Pryhuber from the University of Rochester Medical Center
- For Frozen tissue, rinsed slides for 2X in PBS, then Incubated slides for 5 min in 4% PFA/PBS, then rinsed slides 1X in PBS.
- Rinsed slides for 3 minutes in running dH2O.
- Slides were then incubated in 3% Glacial Acetic Acid (S101, Poly Scientific) for 3 minutes.
- Slides were then incubated in filtered Alcian Blue (S111A, Poly Scientific) for 30 minutes.
- Wash slides under cold running tap water for 10 minutes.
- Incubate for 2 minutes in Nuclear Fast Red (S248, Poly Scientific).
- Wash under flowing dH2O until excess Nuclear Fast Red is washed away.
- Incubated slides in 90% ETOH for 20 dips, repeat in different 90% ETOH.
- Incubated slides in 100% ETOH for 20 dips, repeat in different 100% ETOH.
- Washed slides in 2X in Xylene, followed by 6 more washes in xylene.
- Coverslip with Permount. (Permount Toulene Solution UN 1294, Cat # SP-15-500 Fisher Scientific).
Age: 11 Months