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Alcian Blue Staining of 14 Month-Old Human Lung from Donor D072-RLL-12B2

Donor Tissue Kindly Provided by Dr. Gloria Pryhuber from the University of Rochester Medical Center

Pryhuber_Lab

  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 for 3 minutes in running dH2O.
  4. Slides were then incubated in 3% Glacial Acetic Acid (S101, Poly Scientific) for 3 minutes.
  5. Slides were then incubated in filtered Alcian Blue (S111A, Poly Scientific) for 30 minutes.
  6. Wash slides under cold running tap water for 10 minutes.
  7. Incubate for 2 minutes in Nuclear Fast Red (S248, Poly Scientific).
  8. Wash under flowing dH2O until excess Nuclear Fast Red is washed away.
  9. Incubated slides in 90% ETOH for 20 dips, repeat in different 90% ETOH.
  10. Incubated slides in 100% ETOH for 20 dips, repeat in different 100% ETOH.
  11. Washed slides in 2X in Xylene, followed by 6 more washes in xylene.
  12. Coverslip with Permount. (Permount Toulene Solution UN 1294, Cat # SP-15-500 Fisher Scientific).

 

Tissue Used:
LMH-18-D072-RLL-12B2.15
Gender: Female
Age: 14 Months

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