1 / 26

Introduction to Special Stains

Introduction to Special Stains. Dr Vivien Rolfe De Montfort University This is an Open Educational Resource (OER) that is globally available on the web Creative Commons BY SA. Summary. Why are special stains still important and relevant today?

edan-odonnell
Download Presentation

Introduction to Special Stains

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Introduction toSpecial Stains Dr Vivien RolfeDe Montfort University This is an OpenEducational Resource(OER) that is globally availableon the web Creative Commons BY SA

  2. Summary Why are special stains still important and relevant today? What are some of the chemical principles behind these stains? Some common examples that can be prepared in student laboratory teaching.

  3. What are “special stains”? • H&E was first introduced in the 1870’s and the term special stain came to refer to any technique other than H&E used in the clinical environment. • Whilst the H&E stain is the most common staining method in hospital and research laboratories, it isn’t without its limitations. • H&E cannot visualize micro-organisms. • H&E is not good for distinguishing connective tissue and nerve tissue. • H&E cannot distinguish molecular basis of disease and immunohistochemistry might be preferred.

  4. Six Special Stains!

  5. 1) Toluidine Blue Tolonium Chloride Useful blue cationic dye Cheap and simple application

  6. Neutrophilsseparated from blood sampleStain for 3 minutes in 0.5% toluidine blue

  7. Buccalcells

  8. Distinguishing nerve tissue

  9. 2) Luxol Fast Blue Immunohistochemical methods have advanced but are costly and reagents deteriorate quickly. Special stains include silver methods (such as Gordon and Sweet’s), gold, or Luxol fast blues to stain myelin. Other special stains identify nerve cells. The techniques are important for looking and neurodegeneration.

  10. Distinguishing connective tissueReticulinCollagenFibrin

  11. 3) Gordon and Sweet’s One of several silver methods for staining reticulin. Tissue treated with potassium permanganate to enable the silver to bind. Uses an ammoniacal silver solution. What is reticulin? Why is it important?

  12. Gordon and Sweet’s (Low Power) Liver tissue with no counterstain Reticulin = black

  13. Gordon and Sweet’s (High power) Liver counterstained with what? Reticulin = black Cytoplasm = pink

  14. 4) Van Gieson Trichrome stain – producing 3 colours. Anionic dye and a cationic counterstain. Nuclear stain applied first such as Weigert’shaematoxlin. Collagen stains red with acid fuchsine. Cytoplasm including muscle stains yellow. Washing in acidified water differentiates tissue producing two colours.

  15. Van Giesen (High power) Bladder Collagen = red All other tissue including transitional epithelium = yellow

  16. Van Giesen (Low power) Collagen = red Smooth muscle = yellow Epithelium = yellow Application? Might be used to localisetumours in the bladder to either the smooth muscle or connective tissue layers.

  17. 5) Martius scarlet blue stain Trichrome stain. Martiusyellow and phosphotungstic acid. Brilliant crystal scarlet. Methyl blue. What do the dyes stain?

  18. Tongue (High power) Epithelium = red Collagen = blue Cytoplasm = red No visible yellow

  19. Bladder (low power) Collagen = blue Erythrocytes and early fibrin = yellow Cytoplasm = pink

  20. Adrenal gland (100x oil immersion) Erythrocytes clearly yellow Collagen = blue Cytoplasm = red

  21. Adrenal gland (x100 oil immersion) Early fibrin deposits = diffuse yellow staining Collagen = blue Glandular tissue = red

  22. 6) Masson’s Trichrome Trichrome stain. Iron-haematoxylin plus two anionic dyes. MSB is a variation of this. Iron-haematoxylin. Scarlet-acid fuchsine. Light green (more of a turquoise stain).

  23. Bladder MSB (Left), Masson’s (Right) Nuclei = black Cytoplasm including muscle and epithelium = red Erythrocytes = red Collagen = bluey green or turquoise

  24. Tongue (low power) MSBCollagen = bluey green Cytoplasm of epithelium and skeletal muscle = red

  25. Summary

  26. Further Resources and OERs Histological and Histochemical Methods. 4th Edition. By JA Kiernan. 2007. Scion publishing. Available: http://www.scionpublishing.com Histopathology: Fundamentals of Biomedical Science. By G Orchard and B Nation. 2012. Oxford University Press. Available: http://www.oup.com/uk/orc/bin/fbs/ Laboratory skills open educational resources. De Montfort University. Available: https://www.youtube.com/user/biologycourses

More Related