Frequently asked questions for histochemistry and immunohistochemistry--from John Kiernan's

Table of Contents.

 

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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Protocols / Immunology-immunohistology

Preparing silanized (+plus) slides

Microscopy Techniques
Electron microscopy
HE staining 
Nucleic acid stain
Special cell & cell fraction stains
Antibody purification
Antibody storage and handling
Conjugation of monoclonal antibodies 
Antigen retrieval
Elisa
FRQs for histochemistry
Immunoperoxidase staining techniques
Immunofluorohistochemistry  
Confocal microscope technique 
Laser Capture Microdissection
Immunoprecipitation
ChIP assay

Hybridization in situ

Histotechnology--technical methods

Flow cytometry (FCM)

Kinase assay
 

 

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