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Elastic Fiber Pictures

Human skin VVG 20x
Human skin VVG 20x
Human skin VVG 10x
Human skin VVG 10x

Here are the other 2 pictures that were going to be in the previous post, plus 6 more.  The other 6 are also 10x,20x, and 40x of 2 different tissues.  I can not imagine what life would be like without flexibility.

Aorta Verhoeff Van Gieson 10x
Aorta Verhoeff Van Gieson 10x
Aorta Verhoeff Van Gieson 20x
Aorta Verhoeff Van Gieson 20x
Aorta Verhoeff Van Gieson 40x
Aorta Verhoeff Van Gieson 40x
Human Uterus Verhoeff Van Gieson 10x
Human Uterus Verhoeff Van Gieson 10x
Human Uterus Verhoeff Van Gieson 20x
Human Uterus Verhoeff Van Gieson 20x
Human Uterus Verhoeff Van Gieson 40x
Human Uterus Verhoeff Van Gieson 40x
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Elastic Fibers in the Human Body

Elastic Fibers in the Human Body

I feel like histology is not only a professional field of study but an art form. It takes a series of different arts to make Human skin look like rope strands holding everything together. Elastic fibers help all different parts of the body bend and flex without breaking. Why then do we age? Could we do quantitative and qualitative tests on elastic fibers at different stages of life? They have so many functions and are found in so many areas in and around the body. I have questions about what elastic fibers look like in people who are 40 verses 10 and 80. Are there more or less? Are they thicker, denser, softer or harder. Do they dry out, become brittle or turn into scar tissue when damaged. I’m sure there are researchers who are in the middle of answering many of these questions. Maybe a pathologist already knows. What do you think?
These pictures are the same skin tissue taken at different powers 10x, 20x, 40x. The elastic fibers are the black thread like structures.

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Hematoxylin in Histology

Epididymis H&E 20xHematoxylin, the most commonly used nuclear dye, most commonly used natural dye, extracted from heartwood of the logwood tree which is native to Central America, made in USA! 

Considering all of its versatility in routine and rare variations of Hematoxylin staining, it can stain the following;

Nuclei, mitotic structure, mitochondria, mucin, hemoglobin, elastic fibers, muscle, collagen, axons, phospholipids, protozoa, fatty acids, myelin sheath, alpha and beta cells of the pituitary, pancreatic islets and also certain types of metal.

Oxidants and Mordents-

Hematein is the oxidation product of hematoxylin.

The conversion of hematoxylin to hematein is a process known as ripening which may be achieved naturally through exposure to air (Delafields hematoxylin).

There are also chemical oxidants to hasten the ripening process, mercuric oxide, sodium iodate and potassium permanganate.

PH will have effect on rate of oxidation.

A neutral aqueous solution of hematoxylin will form hematein in a few hours.

Alkaline solutions will affect for a more rapid oxidizing process.

Acid solutions will affect for a more slow oxidizing process.

Mordents- act as a link from the dye to the tissue. Aluminum and Iron are most commonly used, in some cases the mordant is also the oxidizer. For a stable solution the mordant chosen must be a non oxidizer, such as ammonium alum, phosphotungstic acid and phosphomolybdic acid. For a rapidly oxidizing short-lived and unstable solutions that only last up to 24 hours, mordents such as ferric chloride in Wiegerts hematoxylin and also ferric acetate or ferric alum are used.

The combination of mordant and Dye is called a “lake”, in the hematein mordant combination lake there is a positive charge that functions as a cationic or basic dye, this is sometimes called a basophilic stain.

MORE TO COME, READ ABOUT EOSIN AND THE COMBINATION

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Orcein for Elastic Fibers Stain Protocol

This elastic stain is far easier than Verhoeff Van Gieson stain but the chemical can be more unusual to find.

Run a control slide.             Control Tissue – Aorta, Skin

Safety equipment: Work under a hood with lab coat, gloves, and glasses.

  1. De-paraffin slides in xylene (1) —————– 2 minutes (re-use)
  2. De-paraffin slides in xylene (2)—————— 2 minutes (re-use)
  3. Clear slides in 100% alcohol——————— 2 minutes
  4. Clear slides in 100% alcohol——————— 2 minutes
  5. Hydrate slides in 95% alcohol——————- 2 minutes
  6. Hydrate slides in running water—————– 5 minutes
  7. Stain in Orcein ————————————– 30 minutes (re-use)
  8. 70% alcohol —————————————– 10 minutes
  9. 70% alcohol —————————————– 10 minutes
  10. 10.  70% alcohol —————————————- 10 minutes
  11. 11.  Rinse in running water ————————– 1 minute
  12. 12.  Stain in Harris Hematoxylin ——————– 1 minute (re-use)
  13. 13.  Rinse in running water ————————– 1 minute
  14. 14.  Acid alcohol —————————————- 1 dip (re-use)
  15. 15.  Rinse in running water ————————– 5 minutes
  16. 16.  Dehydrate in 95% alcohol———————- 30 seconds
  17. 17.   Dehydrate in 100% alcohol——————- 1 minute
  18. 18.   Dehydrate in 100% alcohol——————- 2 minutes
  19. 19.   Dehydrate in 100% alcohol——————- 2 minutes
  20. 20.   Clear in xylene (3)—————————— 2 minutes (re-use)
  21. 21.   Clear in xylene (4)—————————— 5 minutes (re-use)

Results:         Elastic Fibers – Dark Pink

 

 

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