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Masson’s Trichrome Green Protocol

Masson’s Trichrome Green Protocol

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. Bouin’s Fixative (60 Celsius)———————- 30 minutes (re-use)
8. Wash in running water—————————— 5 minutes
9. Weigerts——————————————- 10 minutes (dump)
10. Wash in running water—————————— 5 minutes
11. Biebrich scarlet acid fuchsin———————- 5 minutes (re-use)
12. Rinse quickly————————————– 30-60 seconds
13. Phostung-Phosmoly acid (PTA-PMA)——————- 5 minutes (re-use)
14. No rinse
15. 2.5% Fast green———————————— 5 minutes (re-use)
16. Rinse quickly ————————————- 30 seconds
17. 1% Glacial acetic acid—————————– 2 minutes (re-use)
18. Dehydrate in 95% alcohol————————— 30 seconds
19. Dehydrate in 100% alcohol————————– 1 minute
20. Dehydrate in 100% alcohol————————– 2 minutes
21. Dehydrate in 100% alcohol————————– 2 minutes
22. Clear in xylene (3)——————————– 2 minutes (re-use)
23. Clear in xylene (4)——————————– 5 minutes (re-use)

Results:
Nuclei——————— Black to purple
Cytoplasm- —————- Shades of pink, red
Erythrocytes and myelin—- Scarlet
Collagen fibers———— Strong bluish-green
Mucus———————- Lighter green

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Histology service

histology service lipid

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Glycosaminoglycans (GAGs)

What are the components of connective tissue?

They are ground substance, extracellular fibers and cellular elements. Ground substance is a matrix that allows diffusion and binds fibers and other tissue elements together.

Components of ground substance are glycoaminoglycans (GAGS), water, calcium.

4 functions of the ground substance are glue, lubrication, barrier, tensil strength.

What are the 2 groups of ground substance,  glycoaminoglycans (GAGS) and glycoproteins.

What are GAGS, they are structural proteins bound to protein core to create a bottle brush structure.

2 types of GAGS are hyaluronic acid and he sulfates.

They have a highly negative because of the sulfate group.

Explain how hyaluronic acid differs from the rest of the GAGS.

Longer chain and forms the base of the bottle brush that the core protein binds to.

Describe a proteoglycan monomer.

A bunch of GAGS around a central protein linked to a hyaluronic acid.

Components of ground substance are: sulfated glycoaminoglycans; heparin sulfate, dermatan sulfate, keratin sulfate, chondroitin sulfate (GAGS sulfated).

What are the function of gags.

They electrostatically bind to collagen for strength, trap water to make a gel that enables tissues to withstand compression.

They are a matrix for cells and blood vessels and a diffusion medium.

http://quizlet.com/9343672/histology-connective-tissue-2-flash-cards/

http://quizlet.com/13755872/histology-lecture-5-packet-2-page-1-19-flash-cards/

There are 5 electrolyte  molar concentrations to stain for. This week I stained for 3 of them. The alcian blue stain works well to detect it.

Every concentration had different results.
0.06 – Carboxylated and sulfated mucosubstances and mucins.
0.3 – Weakly and strongly sulfated mucosubstances and mucins.
0.5 – Strongly sulfated mucosubstances and mucins.
0.7 – Highly sulfated connective tissue mucosubstances.
0.9 Keratan sulfate.

Here are the results:

Rat colon 0.5 10x
Rat colon 0.5 10x
Rat Colon AB 0.7 10x
Rat Colon AB 0.7 10x
Rat colon AB 0.9 10x
Rat colon AB 0.9 10x
Rat knee joint AB 0.5
Rat knee joint AB 0.5