[Histonet] doing Sudan Black B

[John Kiernan]

Hydrated paraffin sections of formaldehyde-fixed tissue are stained the same way as frozen sections of formaldehyde-fixed tissue, but in paraffin sections lipofuscin inclusions are just about the only things that stain. They are, of course, brown or yellow without any staining. If you need to identify lipofuscin inclusions with certainty you'll have to do a few other stains and histochemical tests to distinguish them from ceroid (a similar glycolipoprotein pigment), melanin and haemosiderin.

It is not a good idea to use a protocol informally passed on from someone else without checking its origins. Even a method printed in a book can contain errors, and for this reason just about all the methods books published since about 1950 have references to peer-reviewed papers that readers can and sometimes should check. Any book in our field is just a big collection of review articles that have not been reviewed as critically as more specialized reviews in a good journal.

The late RD Lillie made important original contributions to pigment histochemistry (two are cited below) and his techniques book (also cited) intelligently reviews the subject and provides technical instructions. So does Pearse's Histochemistry.

There are, of course, protocols for staining lipofuscin in all the ordinary histotechniques books. You don't need to have all the latest editions on your lab's bookshelf. The 3rd (1965) edition of Lillie is available for only $8 at https://www.abebooks.co.uk/book-search/title/histopathologic-technic-and-practical-histochemistry/author/r-d-lillie/
Histopathologic Technic and Practical Histochemistry by R D Lillie - AbeBooks<https://www.abebooks.co.uk/book-search/title/histopathologic-technic-and-practical-histochemistry/author/r-d-lillie/>
Histopathologic Technic and Practical Histochemistry by Lillie, R D and a great selection of related books, art and collectibles available now at AbeBooks.co.uk.
www.abebooks.co.uk
Snap it up! The 4th and last (1976) edition of Lillie is now $700 from Amazon, but the 1965 edition is still OK for most of what the book is about, and it's easier to read than his rather rambling 4th edition.

Lillie RD, Fullmer HM (1976) Histopathologic Technic and Practical Histochemistry. 4th ed. McGraw-Hill, New York. See comments above - the previous edition is cheaper and for you it may be just as good.

Pearse AGE, Stoward PJ (1980,1985,1991) Histochemistry, Theoretical and Applied, 4th edn. Vol. 1. Preparative and Optical Technology. Vol. 2. Analytical Technique. Vol. 3. Enzyme Histochemistry. v. 1,2,3. Churchill-Livingstone, Edinburgh.  Pigments are in Vol. 2. Third edn (1968. 2 vols) probably just as good for most labs.

Lillie RD (1956) The mechanism of Nile blue staining of lipofuscins. J. Histochem. Cytochem. 4:377-381. I think all the papers in this journal are now free for anyone to download.

Lillie RD (1956) A Nile blue staining technic for the differentiation of melanin and lipofuscin. Stain Technol. 31:151-153. Sadly, not free unless you have access to a subscribing library or are a member of the Biological Stain Commission.

I hope this answer helps.
John Kiernan
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________________________________
From: LEROY H BROWN via Histonet <histonet at lists.utsouthwestern.edu>
Sent: May 9, 2021 10:29 PM
To: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: Re: [Histonet] doing Sudan Black B

Hi,  I am looking for a protocol for Sudan Black B staining on paraffin
embedded tissue.   Does anyone have a working stain for this?

Thanks

LeRoy Brown HT(ASCP) HTL
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