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

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

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-14B2.3
Gender: Female
Age: 14 Months

By |2019-09-11T12:31:32+00:00October 16th, 2018|Categories: 14 Month-Old, Alcian Blue, Postnatal Human Lung|0 Comments
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Alcian Blue Staining of 14 Month-Old Human Lung from Donor D072-LLL-13B4

Alcian Blue Staining of 14 Month-Old Human Lung from Donor D072-LLL-13B4

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-D072-LLL-13B4.1
Gender: Female
Age: 14 Months

By |2019-09-11T12:30:50+00:00October 15th, 2018|Categories: 14 Month-Old, Alcian Blue, Postnatal Human Lung|0 Comments
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Alcian Blue Staining of 14 Month-Old Human Lung from Donor D072-RLL-15B3

Alcian Blue Staining of 14 Month-Old Human Lung from Donor D072-RLL-15B3

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-15B3.16
Gender: Female
Age: 14 Months

By |2019-09-11T12:29:43+00:00October 15th, 2018|Categories: 14 Month-Old, Alcian Blue, Postnatal Human Lung|0 Comments
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Alcian Blue Staining of 14 Month-Old Human Lung from Donor D072-RLL-12B2

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

By |2019-09-11T12:28:54+00:00October 15th, 2018|Categories: 14 Month-Old, Alcian Blue, Postnatal Human Lung|0 Comments
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Alcian Blue Staining of 14 Month-Old Human Lung from Donor D072-RLL-13B2

Alcian Blue Staining of 14 Month-Old Human Lung from Donor D072-RLL-13B2

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-13B2.17
Gender: Female
Age: 14 Months

By |2019-09-11T12:28:14+00:00October 12th, 2018|Categories: 14 Month-Old, Alcian Blue|0 Comments
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H&E Staining of 14 Month-Old Human Lung from Donor D072-RLL-12B2

H&E 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 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:
LMH-18-D072-RLL-12B2.20
Gender: Female
Age: 14 Months

By |2019-09-11T12:27:01+00:00October 8th, 2018|Categories: 14 Month-Old, H&E, Postnatal Human Lung|0 Comments
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H&E Staining of 14 Month-Old Human Lung from Donor D072-RLL-14B2

H&E Staining of 14 Month-Old Human Lung from Donor D072-RLL-14B2

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 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:
LMH-18-D072-RLL-14B2.17
Gender: Female
Age: 14 Months

By |2019-09-06T19:16:36+00:00October 2nd, 2018|Categories: 14 Month-Old, H&E|0 Comments
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