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60 Cards in this Set

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What are the types of Mucin stains
1) Neutral - found in glands of the GI tract and in prostate. They stain with PAS
2) Acid (simple, or non-sulfated) - Are the typical mucins of epithelial cells containing sialic acid. They stain with PAS, Alcin blue at pH 2.5, colloidal iron, and metachromatic dyes. They resist hyaluronidase digestion.
3) Acid (simple, mesenchymal) - These contain hyaluronic acid and are found in tissue stroma. Stain with Alcian blue at pH 2.5, colloidal iron, and metachromatic dyes. They digest with hyaluronic acid. Found in sarcomas.
4) Acid (complex, or sulfated, epithelial) - These are found in adenocarcinomas. PAS is usually positive. Alcian blue positive at pH 1, colloidal iron, mucicarmine, and metachromatic stains are positive. They resist digestion with hyaluronidase.
5) Acid (complex, connective tissue) - Found in tissue stroma, cartilage, and bone and include substances such as chondroitin sulfate or keratan sulfate. Stain selectively with Alcian blue at pH 0.5.
What are the variety of stains for mucin?
1) Colloidal iron ("AMP")
2) Alcian blue
3) PAS - Stains glycogen as well as mucins,
4) Mucicarmine - V. specific for epithelial mucins.
What are traditional classification of the staining patterns of biogenic amines
1) Chromaffin - have cytoplasmic granules that appear brown when fixed with a dichromate solution - medulla or extraadrenal paraganglion tissues (pheochromocytomas)

2) Argentaffin - reduce a silver solution to metallic silver after formalin fixation - carcinoid tumors of the gut

3) Argyrophil (pre-reduction step necessary)
Types of stains for argentaffin include:
1. Diazo (diazonium salts)
2. Fontana-Masson
3. Schmorl's
4. Autofluorescence
Types of stains for chromaffin include:
1. Modified Giemsa
2. Schmorl's
3. Wiesel's
Types of stains for argyrophil include:
1. Grimelius (Bouin's fixative preferred)
2. Pascual's
Where is melanin found
Found in the skin, eye, and substantia nigra. It may also be found in melanomas.
What are the types of melanin stains
Fontana-Masson

Schmorl's method

DOPA-oxidase

Formaldehyde-induced fluorescence
What is the Fontana-Masson
Relies upon the melanin granules to reduce ammoniacal silver nitrate (but argentaffin, chromaffin, and some lipochrome pigments also will stain black as well).
How does Schmorl's method work
Uses the reducing properties of melanin to stain granules blue-green.
What is the most specific method of melanin staining
DOPA-oxidase
How does DOPA-oxidase work
Requires frozen sections for best results, but paraffin sections of well-fixed tissues may be used.
The stain works because the DOPA substrate is acted upon by DOPA-oxidase in the melanin-producing cells to produce a brownish black deposit.
What technique removes melanin
Bleaching techniques - make use of a strong oxidizing agent such as potassium permanganate or hydrogen peroxide.
What does pseudomelanin stain with
PAS
True melanin isn't PAS positive
What are Lipochrome (lipofuschin) pigments?
breakdown products within cells from oxidation of lipids and lipoproteins
Where are lipochrome/lipofuschin pigments commonly found
most commonly in heart, liver, CNS, and adrenal cortex (zona reticularis).
What are ceroid pigments and where are they found?
Less highly oxidized lipochrome pigments found in the testis interstitium and seminal vesicle
Where is hemosiderin found
in areas of old hemorrhage or be deposited in tissues with iron overload
What method is used to stain iron tissues
Perl's iron stain
How is Perl's iron stain produced?
The section is treated with dilute hydrochloric acid to release ferric ions from binding proteins.
Then these ions then react with potassium ferrocyanide to produce an insoluble blue compound (the Prussian blue reaction)
What methods are used for calcium staining
Hematoxylin - blue color

VonKossa stain - silver reduction

Alizarin red S - orange-red lake

Azan - differentiates osteoid from mineralized bone
When is a VonKossa stain used?
Demonstrates phosphates and carbonates, but these are usually present along with calcium.

Most useful when large amounts are present, as in bone.
When is Alizarin red S used?
at pH = 4.2
It works best with small amounts of calcium (such as in Michaelis-Gutman bodies).
Where are urates present in the body
in tissues at sodium urates
How are urates fixed
With 95% or absolute alcohol because they are soluble in aq. solns
What stains are for urates
Methenamine silver - stains urates black.
What is a special property of urates
birefringent on polarization

Using a red plate, the crystals show negative birefringence (yellow color) when the crystal's long axis is aligned in the direction of the slow wave. At 90 degrees to this, the crystals will be blue.
What stains are used for copper
rubeanic acid and rhodanine stains are utilized to detect the cytoplasmic accumulation of copper in the liver.
What is Wilson's disease
autosomal recessive disease resulting from decreased serum ceruloplasmin, the blood protein that transports serum copper
What are examples of exogenous pigments and minerals
carbon, asbestos, silica, street drugs, tatto pigments,
How does carbon appear and how is it imaged?
- appears as anthracotic pigment in the lungs
- can be distinguished from melanin by doing a melanin bleach
-
What characterizes asbestos in tissue?
these crystals are highly irritative and highly fibrogenic. The fibers become coated with a protein-iron-calcium matrix, giving them a shish-kebab appearance.
What is used to stain asbestos?
an iron stain
How is silica visualized?
It is inert and can't be stained

It's demonstrated by white birefringence on polarization
What characterized tattoo pigment
black, inert, non-polarizable.

Red tattoo pigment often contains cinnabar (which has mercury in it).
How are minerals visualized?
In general, minerals are best demonstrated by microincineration techniques or by scanning electron microscopy with energy dispersive analysis (SEM-EDA).
How is fat stained?
oil red O (ORO) stain can identify neutral lipids and fatty acids in smears and tissues
Why are fresh smears or cryostat sections of tissue necessary in fat stains
because fixatives containing alcohols, or routine tissue processing with clearing, will remove lipids
What stains are used to visualize CT?
1) trichrome stain
2) reticulin stain
3) elastic tissue stain
What is an elastic tissue stain used for?
helps to outline arteries, because the elastic lamina of muscular arteries, and the media of the aorta, contain elastic fibers.
What stains ID inflammatory cells in peripheral blood and tissue
1) Wright-Giemas and Giemsa stains
2) leukocyte alkaline phosphatase (LAP)
3) tartrate-resistant acid phosphatase (TRAP)
4) myeloperoxidase (MPO).
What is a special property of giemsa stains
One property of methylene blue and toluidine blue dyes is metachromasia. This means that a tissue component stains a different color than the dye itself. For example, mast cell graules, cartilage, mucin, and amyloid will stain purple and not blue, which is helpful in identifying these components.
What is a LAP stain used for
- in determining whether a high peripheral blood leukocytosis is a reactive process or a leukemia (chronic myelogenous leukemia, or CML).
- more differentiated cells in the reactive process will stain more readily with LAP, while leukemic cells will not
What does a high LAP score indicate
high score generally indicates a "leukemoid reaction" or reactive condition (with an infection or other inflammatory process)
What does a low LAP score indicate
a low score suggests CML (leukemia)
What is the use of a TRAP stain
help diagnose a rare leukemia known as hairy cell leukemia.
What is an MPO stain used for
to identify cytoplasmic granules characteristic of myeloid cells - helps differentiate between myeloid and lymphoid cells
What is used to stain bacteria
H&E
How does bacteria appear upon staining
Bacteria appear on H and E as blue rods or cocci regardless of gram reaction. Colonies appear as fuzzy blue clusters.
What is used to stain gram(-) bac.
Brown and Brenn (or the Brown and Hopps modification)
What stain fungi
H&E: blue
PAS: red
Methenamine silver: most sensitive
What demonstrates donovan bodies and leishmania?
Giemsa stain
What is a Warthin-Starry stain used for
to ID Spirochetes - but they are difficult to ID
What is an AFB (acid fast bacilli) stain used to
uses carbol-fuchsin to stain the lipid walls of acid fast organisms such as M. tuberculosis, myobacteria, cryptosporidium, isospora, and the hooklets of cysticerci.
What are the methods of AFB stains
1) Ziehl-Neelsen method
2) Kinyoun's method
3) Fite stain
4) auramine stain - most sensitive, req. fluorescence microscope
What is a Gomori methenamine silver (GMS) stain used for
fungi and for Pneumocystis carinii (jirovecii) - organisms are outlined by the brown to black stain
What stains fungi and Pneumocystis carinii (jirovecii)
GMS
What does a PAS stain visualize?
glycogen, mucin, mucoprotein, glycoprotein, as well as fungi
What is a PAS stain useful for
outlining tissue structures--basement membranes, capsules, blood vessels, etc.
What stains can have a high background
GMS, PAS