Posted on Leave a comment

Alizarin Red S Stain Protocol for Calcium

Run a control slide. Control tissue: Bone or tissue with calcium present

Safety equipment: Work under a hood with lab coat, gloves, and safety 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 50% alcohol——————– 1 minute
  7.   Rinse rapidly in DH2O—————————— 30 seconds
  8.   Cover the sections with alizarin Red S solution (re-use)
  9.   Observe the reaction under a microscope and remove when the red-orange lake forms ( 5 minutes to 15 mintues). The lake should be heavy but not too diffuse.

10.    Shake off excess dye and blot carefully.

11.    Dehydrate in acetone for 10 to 20 seconds (dump)

12.    Dehydrate in acetone-xylene (50:50)———–10 to 20 seconds (dump)

13.    Clear in xylene—————————————- 5 minutes (re-use)

14.    Coverslip

Results:

Calcium Deposits (except oxalate) ———————Orange-Red

This precipitate is birefringent.

 

An Expert Histology Service

Posted on Leave a comment

Hematoxylin & Eosin (H&E) Protocol

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 slides in Harris Hematoxylin————–  10 minutes (re-use)
  8. Rinse slides in running water———————  2 minutes
  9. Differentiate in acid alcohol———————-   1 quick dip (re-use)
  10. Rinse in water————————————–    3 dips
  11. Dip in ammonia water—————————-    15-30 seconds (re-use)
  12. Wash in running tap water———————–    10 minutes
  13. Place in 95% alcohol——————————    2 minutes
  14. Stain in eosin—————————————    1 minute (re-use)
  15. Dehydrate in 95% alcohol————————   1 minute
  16. Dehydrate in 100% alcohol———————-   1 minute
  17. Dehydrate in 100% alcohol———————-   2 minutes
  18. Dehydrate in 100% alcohol———————-   2 minutes
  19. Clear in xylene (3)———————————   2 minutes (re-use)
  20. Clear in xylene (4)———————————   5 minutes (re-use)

Results: Nuclei –Blue   Cytoplasm – Pink     Blood – Red

Posted on 3 Comments

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

Posted on Leave a comment

Histology service

histology service lipid

We have an extensive array of histology knowledge, from years of working in clinical, research and academia histology labs.  Our techniques have been modified to give you excellent results and turnaround times.  The desire to help your causes, through research advance faster through the end stage of a histology service drives our success. We understand that your research is the most important thing in the world and our job is to make your life easy by doing everything right on our end within a reasonable time.

Do to this, we have built our lab for efficiency and quality.  All specimens come in and leave in a circular fashion. This eliminates redundancy and waste by creating checks and balances that make a lean lab.  We believe, it’s our job to make you look good.

Come see what we can do for you.