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Alcian Blue Staining of 11 Month-Old Human Lung from Donor D094-LLL-5B2

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

Pryhuber_Lab

 

  1. 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.
  2. Rinsed slides for 3 minutes in running dH2O.
  3. Slides were then incubated in 3% Glacial Acetic Acid (S101, Poly Scientific) for 3 minutes.
  4. Slides were then incubated in filtered Alcian Blue (S111A, Poly Scientific) for 30 minutes.
  5. Wash slides under cold running tap water for 10 minutes.
  6. Incubate for 2 minutes in Nuclear Fast Red (S248, Poly Scientific).
  7. Wash under flowing dH2O until excess Nuclear Fast Red is washed away.
  8. Incubated slides in 90% ETOH for 20 dips, repeat in different 90% ETOH.
  9. Incubated slides in 100% ETOH for 20 dips, repeat in different 100% ETOH.
  10. Washed slides in 2X in Xylene, followed by 6 more washes in xylene.
  11. Coverslip with Permount. (Permount Toulene Solution UN 1294, Cat # SP-15-500 Fisher Scientific).

 

 

Tissue Used:
LMH-18-D094-LLL-5B2.4
Gender: Male
Age: 11 Months