Staining,
Histochemistry and Histotechnology
FAQ (Frequently Asked Questions)
Version 1.3 January 2007. (Version 2.0 is in the works, 2008.)
Compiled
by: J.A. Kiernan,
Department of Anatomy and Cell
Biology,
The University of Western Ontario
London, Canada
Link to the Biological Stain Commission
TABLE
OF CONTENTS
Introduction
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 internet has
been a valuable source of questions and answers, and I am grateful to
many people 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 within a few days.
These FAQ items do not have the authority of peer-reviewed scientific publications.
The BSC's bimonthly journal, Biotechnic & Histochemistry, has a Notes and Queries
feature with expert-reviewed answers to questions
about histochemistry, histology, histotechnology,immunostaining, traditional stains etc. Notes and Queries items are peer-reviewed; the authors of these items receive frequent email requests for PDF reprints.
The new version of the FAQ for the BSC web site is not yet ready (October 2008).
For a recent (2007) version, click this link to the compiler's web site. Use your
browser's back-button to return to this page of the BSC site.
= = =
The liinks below are experimental. Ignore them because most go nowhere.
FIXATION,
FREEZING ETC
**
Carbodiimides as fixatives
**
Carnoy & alcoholic fixatives
**
Perfusion fixative for electron microscopy
**
Fixation of frozen sections.
**
Non-formaldehyde commercial fixatives
**
Glutaraldehyde and immunohistochemistry
**
Isopentane: alternative names
**
Lidocaine in perfusion fixation
**
Michel's fluid for transporting cells or specimens
**
Microwave ovens: Advice for new users
**
Paraformaldehyde: why won't it dissolve?
**
Saccomano's fixative
**
Zinc-containing fixatives: What has been published?
**
Alternatives to mercury-containing fixatives
PROCESSING,
DECALCIFYING, EMBEDDING
**
Solvent to replace xylene AND alcohols
**
2-butoxyethanol ("Clereum") dehydrating or clearing agent
**
Decalcification: Acid or EDTA?
**
Testing for completeness of decalcification
**
Fatty specimens: Processing into paraffin.
**
Polymethyl methacrylate embedding for bone
**
Mold release spray
**
Paraffin processing of skin
**
Cryoprotection of specimens
**
Cutting sections of toe or finger nails
**
Paraffin wax: crystals, additives and cutting
**
Xylene substitutes: what are they?
**
Test for water in used absolute alcohol
**
Molecular sieves for making anhydrous solvent
SECTIONING,
SLIDE ADHESIVES, MOUNTING
**
Sections coming off slides. Which adhesive?
**
Apathy's mounting medium and variants
**
Silanized (APES or TES or positively charged) slides
**
Polishing undecalcified bone sections.
**
Polylysine-coated slides
**
Wrinkles in plastic sections
**
Wrinkles in paraffin sections containing cartilage
**
Thick paraffin sections
**
Sectioning plastic-embedded specimens
**
Iodine for removing mercury deposits
**
Labeling slides
**
Sectioning plant material: some hints.
STAINING
METHODS, HISTOCHEMISTRY
** Making
aldehyde-fuchsine
** Phosphatases
in decalcified, embedded tissue.
** Congo
red for amyloid
** Cartilage
staining with safranine
** Stain
for Chlamydia (Castaneda's method).
** Which
staining method for copper is best?
** Diastase
(amylase) control for glycogen
** Evans
blue, trypan blue and eosin as tracers.
** Gallyas'
stain
** Gram
staining of sections (Brown & Hopps method).
** Oxidants
for hematoxylin
** McFaydean's
stain for anthrax bacilli
** Microglia
with Griffonia lectin.
** Picro-sirius
red staining
** Iron
hematoxylin: ripening not needed.
** Enzyme
histochemistry on cell cultures
** Malachite
green in stain for Cryptosporidium
** Confusing
dye names (lissamine fast red as an example)
** Mayer's
and Gill's hematoxylins
** Effects
of pH on staining by dyes
** Histochemical
stain for arsenic
** Giemsa
staining of blood smears: several hints
** Automated
H & E staining problems
** Verhoeff's
stain for myelin and elastin
** Acridine
orange method for DNA and RNA
** Quickly
finding something in a newly cut section
** Fluorescent
lectins: general method
** Methyl
blue and methylene blue
IMMUNOHISTOCHEMISTRY
** Paraffin
or frozen sections for immunohistochemistry
** Inhibiting
endogenous peroxidase
** Using
mouse primary antibodies on mouse tissues
** Antigen
retrieval: A patented or copyright phrase?
** p53
protein
** Prevention
of fluorescence fading
** Background
in immunostained cartilage
** Endogenous
biotin in mast cells?
MISCELLANEOUS
STUFF
** Disposal
of used diaminobenzidine (DAB) solutions
** Dilution
of concentrated acids: formula etc.
** Disposal
of waste from "special stains"
** Magnification
of a photomicrograph
** Can
a method be both published and patented?
** Books
and articles about artifacts in histology
** How
dangerous is picric acid?
** Which
color print film for photomicrography?
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