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H&E Staining of 31 Year-Old Human Lung from Donor CCHMC-D003-A21.12

H&E Staining of 31 Year-Old Human Lung from Donor CCHMC-D003-A21.12

 

 

The tissue studies were done in collaboration with Francis X. McCormack, MD and Kathryn A. Wikenheiser-Brokamp, MD/PhD with support of LifeCenter and CCHMC Patholgy Research Core

  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 stained in filtered Hematoxylin for 60 seconds. (Gold Standard Harris Hemotoxylin, Acidified, Mercury; Cat # 24245 Polysciences Inc,).
  4. Rinsed slides in tap water until water turned cleared then rinsed in dH2O
  5. 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.
  6. Rinsed slides in dH2O 3X 20 dips.
  7. Rinsed slides in 70% ETOH for 20 dips.
  8. Rinsed slides in 80% ETOH for 20 dips.
  9. Stained slides in Eosin for 15 seconds. (Eosin Y 0.5% Alcohol Solution, Cat # 09859 Polysciences Inc,).
  10. 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.
  11. Rinsed slides 3X in 100% ETOH. 10 dips for the first and second washes and 15 dips in the third and fourth washes.
  12. Rinsed slides in 2X in Xylene, followed by 6 more washes in xylene.
  13. Coverslip with Permount. (Permount Toulene Solution UN 1294, Cat # SP-15-500 Fisher Scientific).

Tissue Used: 
LMH-18-CCHMC-D003-A21.12
Age: 31 Years
Gender: Female

By |2019-09-11T13:15:22+00:00November 2nd, 2018|Categories: 31 Year Old, Adult Human Lung, H&E|0 Comments
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Alcian Blue Staining of 11 Month-Old Human Lung from Donor D094-LLL-4B3

Alcian Blue Staining of 11 Month-Old Human Lung from Donor D094-LLL-4B3

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-4B3.6
Gender: Male
Age: 11 Months

By |2019-09-11T12:23:04+00:00October 19th, 2018|Categories: 11 Month-Old, Alcian Blue, Postnatal Human Lung|0 Comments
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Alcian Blue Staining of 7 Year-Old Human Lung from Donor D078-RLL-18B3

Alcian Blue Staining of 7 Year-Old Human Lung from Donor D078-RLL-18B3

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-D078-RLL-18B3.11
Gender: Female
Age: 7 Years

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By |2018-10-31T16:58:57+00:00October 10th, 2018|Categories: 7 Year-Old, Alcian Blue, Human Lung Disease, Postnatal Human Lung|0 Comments
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Alcian Blue Staining of 7 Year-Old Human Lung from Donor D078-RLL-17B4

Alcian Blue Staining of 7 Year-Old Human Lung from Donor D078-RLL-17B4

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-D078-RLL-17B4.12
Gender: Female
Age: 7 Years

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By |2018-10-31T16:57:12+00:00October 10th, 2018|Categories: 7 Year-Old, Alcian Blue, Human Lung Disease, Postnatal Human Lung|0 Comments
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Alcian Blue Staining of 7 Year-Old Human Lung from Donor D078-RLL-16B2

Alcian Blue Staining of 7 Year-Old Human Lung from Donor D078-RLL-16B2

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-D078-RLL-16B2.13
Gender: Female
Age: 7 Years

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By |2018-10-31T16:55:42+00:00October 10th, 2018|Categories: 7 Year-Old, Alcian Blue, Human Lung Disease, Postnatal Human Lung|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-15B3

H&E 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 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-15B3.18
Gender: Female
Age: 14 Months

By |2019-09-11T12:26:19+00:00October 5th, 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-13B2

H&E 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 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-13B2.19
Gender: Female
Age: 14 Months

By |2019-09-11T12:25:32+00:00October 5th, 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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H&E Staining of 11 Month-Old Human Lung from Donor D094-LLL-4B3

H&E Staining of 11 Month-Old Human Lung from Donor D094-LLL-4B3

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 stained in filtered Hematoxylin for 60 seconds. (Gold Standard Harris Hemotoxylin, Acidified, Mercury; Cat # 24245 Polysciences Inc,).
  4. Rinsed slides in tap water until water turned cleared then rinsed in dH2O
  5. 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.
  6. Rinsed slides in dH2O 3X 20 dips.
  7. Rinsed slides in 70% ETOH for 20 dips.
  8. Rinsed slides in 80% ETOH for 20 dips.
  9. Stained slides in Eosin for 15 seconds. (Eosin Y 0.5% Alcohol Solution, Cat # 09859 Polysciences Inc,).
  10. 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.
  11. Rinsed slides 3X in 100% ETOH. 10 dips for the first and second washes and 15 dips in the third and fourth washes.
  12. Rinsed slides in 2X in Xylene, followed by 6 more washes in xylene.
  13. Coverslip with Permount. (Permount Toulene Solution UN 1294, Cat # SP-15-500 Fisher Scientific).

 

Tissue Used:
LMH-D094-LLL-4B3.14
Gender: Male
Age: 11 Months

By |2019-09-11T12:20:32+00:00September 28th, 2018|Categories: 11 Month-Old, H&E, Postnatal Human Lung|0 Comments
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Alveolar type II cells, also known as type II pneumocytes, are specialized epithelial cells found in the alveoli of the lungs, which are the tiny air sacs responsible for gas exchange. These cells play a crucial role in maintaining the structure and function of the respiratory system.

Alveolar type II cells are located within the alveolar walls of the lungs.

Alveolar type II cells are cuboidal or squamous in shape and are smaller than alveolar type I cells, which are responsible for gas exchange. They are characterized by the presence of microvilli on their apical surface, which increases their surface area for various functions.

One of the most important functions of alveolar type II cells is the production and secretion of pulmonary surfactant. Surfactant is a complex mixture of lipids and proteins that reduces the surface tension of the alveolar fluid, preventing the collapse of alveoli during exhalation. This property is crucial for maintaining the stability of the alveoli and preventing lung collapse, particularly at the end of expiration.

Alveolar type II cells serve as progenitor cells for alveolar type I cells. They have the ability to self-renew and differentiate into type I cells, helping in the repair and regeneration of the alveolar epithelium after injury or damage. This regenerative capacity is essential for maintaining the integrity of the respiratory barrier.

Alveolar type II cells also play a role in the innate immune response of the lungs. They can produce and secrete various immune molecules, including cytokines, chemokines, and antimicrobial peptides, in response to pathogens or inflammatory stimuli. These molecules help to recruit immune cells to the site of infection and promote the clearance of pathogens from the lungs.

Alveolar type II cells are involved in the regulation of ion and fluid balance within the alveoli. They express various ion channels and transporters that regulate the movement of ions, such as sodium and chloride, across the epithelial barrier. This process is important for maintaining the osmotic balance of the alveolar fluid and preventing the accumulation of fluid in the lungs (pulmonary edema).

The surfactant produced by alveolar type II cells consists mainly of phospholipids (such as dipalmitoylphosphatidylcholine, or DPPC) and surfactant proteins (SP-A, SP-B, SP-C, and SP-D). These components work together to reduce surface tension and maintain the stability of the alveoli.

The production and secretion of surfactant by alveolar type II cells are regulated by various factors, including mechanical stretch, glucocorticoid hormones, and certain signaling molecules (such as epinephrine and thyroid hormones). These regulatory mechanisms ensure that surfactant production is matched to the demands of respiration and the maintenance of lung function.