Donor tissue kindly provided by Dr. Scott Randell, University of North Carolina

  1. Paraffin sections were placed at 60oC overnight to melt paraffin.
  2. Paraffin sections were then placed in xylene (Xylenes Reagent Grade ACS Histology/Lab Grade Cat # 399000000 Pharma-AAPER) 3X for 10 minutes each.
  3. Rinsed slides in a series of 100% ETOH washes. 20 dips for the first wash, 3 minutes for the second wash, and 20 dips for the third wash.
  4. Rinsed slides in a series of 95% ETOH washes. 20 dips for the first wash, 3 minutes for the second wash, and 20 dips for the third wash.
  5. Rinsed slides in 70% ETOH for 3 min.
  6. Rinsed slides for 3 minutes in running dH2O.
  7. Slides were then incubated in 3% Glacial Acetic Acid (S101, Poly Scientific) for 3 minutes.
  8. Slides were then incubated in filtered Alcian Blue (S111A, Poly Scientific) for 30 minutes.
  9. Wash slides under cold running tap water for 10 minutes.
  10. Incubate for 2 minutes in Nuclear Fast Red (S248, Poly Scientific).
  11. Wash under flowing dH2O until excess Nuclear Fast Red is washed away.
  12. Incubated slides in 90% ETOH for 20 dips, repeat in different 90% ETOH.
  13. Incubated slides in 100% ETOH for 20 dips, repeat in different 100% ETOH.
  14. Washed slides in 2X in Xylene, followed by 6 more washes in xylene.
  15. Coverslip with Permount. (Permount Toulene Solution UN 1294, Cat # SP-15-500 Fisher Scientific).

Tissue Used:

LMHA-15-UNC-DD036L-04HP-2.12

Gender: Male

Age: 47 Year Old

Race: Caucasian

Non-Smoker