Histology FAQ

Staining, Histochemistry and Histotechnology

(Frequently Asked Questions)

 

Dr. John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London, Canada

 

 

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Introduction

The questions and answers are grouped into six categories:

Fixation, freezing etc Processing, decalcifying, embedding Sectioning, slide adhesives, mounting Staining methods, histochemistry Immunohistochemistry Miscellaneous stuff

In an attempt to cut down on spam, all the email addresses in this document have the @ sign replaced with [AT]. If you send an email to one of these addresses remember to edit the address to restore the @.

The title of each item is followed by a Question (sometimes more than one, if they are closely related), and then by one or more Answers. Items vary in length. Most consist of one or two screenfuls of text. A few topics are differently treated, to achieve a more effective way to answer some questions.

It should be noted that the intention of this FAQ is to explain things, not to provide a compendium of favorite recipes. There are textbooks, and also other web sites, that provide detailed instructions for making solutions and performing techniques. The properly educated technician, pathologist or research worker understands the reason for each step in a procedure. Simple modifications will always be needed to adapt "standard" methods to particular applications. Often, a little study and a lot of thought can shorten the trial and error approach to staining.

Visitors to the web site are encouraged to submit new questions (or, better still, questions with answers) to be considered for inclusion in later releases of this FAQ. Corrections and other suggestions will also be welcome. Questions and comments may be sent to the compiler by email:  <J. A. Kiernan>  kiernan[AT]uwo.ca

Questions and answers in print

Starting in January 2001 the journal Biotechnic & Histochemistry carries a feature called Notes and Queries consisting of anonymous questions with signed, peer-reviewed answers. For controversial issues there may be more than one answer. The peer review process means that a Note responding to a Query in B & H is a statement made in a journal that has had strict editorial standards for more than 75 years. (The name of the journal was changed from Stain Technology to B & H in 1991.)

Biotechnic & Histochemistry is published 6 times a year. Every issue contains interesting papers that emphasize techniques and their intelligent implementation. B & H is the journal of the Biological Stain Commission and it provides good value for money for libraries (about $200) and individual subscribers (about $100; but with 30% off for trainees, and an even better discount if you're a member of the Biological Stain Commission).

For information about B & H click on this link: Biotechnic & Histochemistry
This link: Biological Stain Commission will tell you about the B.S.C. - what it is and what it does.

Information in a peer-reviewed journal such as B & H is usually right, because it has been subjected to a severe reviewing process. Scientists are not nice to one another when doing peer reviews. It's part of our job to ensure that the papers in a journal accurately report work that has been carefully and intelligently carried out before allowing it to be published. Contributions that don't make the grade are rejected.

Acknowledgements

I thank the many people who have answered my questions about staining and related methodology, and also those who have asked my advice and made me think and investigate. In recent years the HistoNet listserver has been a valuable source of questions and answers, and I am grateful to many of its contributors who have kindly allowed me to reproduce their wisdom here. The Questions in this FAQ are all anonymous, but the sources of the Answers are all acknowledged.

Permission was requested and granted for all the Answers provided by people other than myself. This involved much exchanging of emails, which may not always have been received and answered. It is therefore possible that I have erred by including a few Answers without written permission. If, gentle reader, you see yourself quoted without consent in this FAQ, please email me at kiernan[AT]uwo.ca. I will immediately expunge the offending Answer and find or write another to replace it. If you want to revise something attributed to you, let me know and I'll incorporate the change as soon as I get a round tuit.

We are very grateful for Dr. John A. Kiernan generous contribution of this summary article. For more information and latest updates, please visit his website at http://publish.uwo.ca/~jkiernan/