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 stained in filtered Hematoxylin for 60 seconds. (Gold Standard Harris Hemotoxylin, Acidified, Mercury; Cat # 24245 Polysciences Inc,).
  8. Rinsed slides in tap water until water turned cleared then rinsed in dH2O
  9. Rinsed slides in Lithium Carbonate: 20 dips. (Lithium Carbonate ACS Grade; Cat # 255823-500G Sigma Aldrich). Lithium carbonate is prepared by over-saturating in dH2O.
  10. Rinsed slides in dH2O 3X 20 dips.
  11. Rinsed slides in 70% ETOH for 20 dips.
  12. Rinsed slides in 80% ETOH for 20 dips.
  13. Stained slides in Eosin for 15 seconds. (Eosin Y 0.5% Alcohol Solution, Cat # 09859 Polysciences Inc,).
  14. Rinsed slides in a series of 95% ETOH washes. 10 dips for the first and second washes, 15 dips in the third and fourth washes.
  15. Rinsed slides 3X in 100% ETOH. 10 dips for the first and second washes and 15 dips in the third and fourth washes.
  16. Rinsed slides in 2X in Xylene, followed by 6 more washes in xylene.
  17. Coverslip with Permount. (Permount Toulene Solution UN 1294, Cat # SP-15-500 Fisher Scientific).

Tissue Used:

LMHA-UNC-DD034L-03HP-3-57

Gender: Male

Age: 31 Year Old

Race: Hispanic

Non-Smoker