Woodland Pharmaceuticals Metastasis models

Posted by Histologistics on

Hello everyone, we are proud to announce that we have been working with Woodland Pharmaceuticals in their project to identify a liver to lung metastasis model (MHCC97H) and are evaluating the lung cancer SQ A549 for lung lesions as well. At this point Woodland Pharmaceuticals is establishing a stable luciferase to express tumor cell lines with their partner SBH Sciences so metastasis can be quantified. They have positive control compounds that will mechanistically block most/all of this metastasis.  We have done all their histology so far and have much more to do.  Follow us with this exciting project.

Here are some of the pictures.

 

Primary tumor 10x

Primary tumor 10x

 

 

 

 

 

Mouse lung metastasis 40x

Mouse lung metastasis 40x

 

 

 

 

 

 

Liver metastasis 20x

Liver metastasis 20x

 

 

 

 

 

Liver metastasis 40x

Liver metastasis 40x

 

 

 

 

 

 

If you have LinkedIn account, you can take look at this exciting project at;  http://www.linkedin.com/company/2696725?trk=tyah

Alcian Blue / PAS Stain Protocol

Posted by Histologistics on

Here is the protocol for alcian blue periodic acid schiff  (AB /PAS).  At the bottom of the post is the link for printing.

Control tissue: Large Intestine

Rat colon PASAB2

  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. Place slides in 3% Acetic Acid————- 3 minutes (re-use)
  8. Alcian Blue ———————————– 30 minutes (re-use)
  9. Wash in running water ——————— 10 minutes
  10. Oxidize in 0.5% periodic acid ————- 10 minutes (re-use)
  11. Wash in running water ——————— 5 minutes
  12. Schiff reagent (room temp)—————- 20 minutes (re-use)
  13. Wash in running water ——————— 10 minutes
  14. Dehydrate in 95% alcohol—————— 30 seconds
  15. Dehydrate in 100% alcohol—————- 1 minute
  16. Dehydrate in 100% alcohol—————- 2 minutes
  17. Dehydrate in 100% alcohol—————- 2 minutes
  18. Clear in xylene (3)————————— 2 minutes (re-use)
  19. Clear in xylene (4)————————— 5 minutes (re-use)

Results: Polysaccharides and Mucosubstances containing hexoses or deoxyhexoses with vicinal glycol groups – Magenta to Red

Hyaluronic acid, sialomucins and all but the most strongly acidic sulfated mucosubstances – Blue

PAS Alcian Blue

Acid Fast Bacilli (AFB) Protocol

Posted by Histologistics on

Here is the protocol for acid fast bacilli (AFB).  At the bottom of the post is the link for printing.

Control Tissue : Tissue with AFB present

Human Kidney AFB

Human Kidney AFB

  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.   Carbol Fuchsin————————————– 45 minutes (re-use)
  8.   Wash in running water—————————– 1 minute
  9.   Decolorize in 1% acid alcohol——————— until sections are pale pink
  10.   Wash in running water—————————– 8 minutes
  11.   Counterstain with working methylene blue—–1 dip (re-use (good for 5 days))
  12.   Rinse quickly—————————————- 30 seconds
  13.   Dehydrate in 95% alcohol———————— 1 minute
  14.   Dehydrate in 100% alcohol———————- 1 minute
  15.   Dehydrate in 100% alcohol———————- 2 minutes
  16.   Clear in xylene (3)——————————— 2 minutes (re-use)
  17.   Clear in xylene (4)——————————— 5 minutes (re-use)

Results:

Acid-fast bacilli —————- Bright Red

Erythrocytes——————– Yellowish Orange

Other tissue elements——- Pale blue

Acid Fast Bacilli

Luxol Fast Blue Stain Variations

Posted by Histologistics on

We did this as a test to see different versions of luxol fast blue (LFB). LFB is a stain for the myelin sheath or myelinated axons in brain and spinal cord.  We tested on rat brain.  There are 4 variations.  They are; LFB / Cresyl violet (CR), LFB /H&E, LFB / Periodic acid schiff (PAS) and LFB / nuclear fast red (NFR).   The H&E and NFR versions are for background staining only.  The PAS could show glycogen or fungus present and the CR shows nissl bodies.

Rat brain LFB / CV 20x

Rat brain LFB / CV 20x

Rat brain LFB / H&E 20x

Rat brain LFB / H&E 20x

Rat brain LFB / NFR 20x

Rat brain LFB / NFR 20x

Rat brain LFB / PAS 20x

Rat brain LFB / PAS 20x

Histology service

Posted by Histologistics on

We specialize in research projects.  All projects received are completed by the person who receives it.  This eliminates problems created by incomplete or incorrect communication.  On small projects all phases of histology can usually be completed within 2 days of receipt.  During the first day, we log, gross and cassette if necessary, and process.  On the second day, we embed, cut and stain.  To make sure everything is correct, we check every slide against its block and manifest 3 times by someone other than the person who cut them.  Doing this takes a little longer but eliminates most technical errors.  For this reason we require a manifest of all specimens sent before any work is done.  manifest-list-sheet.xlsx