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Hem Yr 2 Sem 2 > Aclian Blue Stain > Flashcards

Aclian Blue Stain Flashcards

1
Q

What is the target of alcian blue

A

Mucin
carbohydrate group
mono - simple sugar
poly-
muco polysacc - long chain of sugars found in the body , needed for cell growth and structure

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2
Q

What is part of group 1 poly sacc

A

-carry no charge
Neutral poly (non ionic homoglycans)
-found in stomach and bruners glands
-Glucose containing glycogen, starch and cellulose
N-acetyl -glucoasmine containing CHITIN

PAS positive

AB negative , Collodial iron negative, Mayers mucicarmine method MMM positive

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3
Q

What is part of carbohydrate group 2

A

Acid mucopolysaccharides (anionic heteroglycans) CARRY A NEGATIVE CHARGE ON MUCIN MOLECULE

Classifed as simple with
Carboxyl group- hyaluronic acid (connect tissue and umbilical cord)

Complex
Sulfuric group and Carboxy : chondroitin A, B, C (cartilage, cornea, blood vessels, aorta) and Heparin

Sulfated : Aorta

All acidic and attached to proteins

Pas NEGATIVE

AB positive
Colloidal iron POSITIVE
MMM Positive

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4
Q

Group 3 carbs consist of

A

Glycoproteins (mucins, mucoid, mucoprotein, mucocutaneus )
mainly epithelial mucins

neutral - stomach mucin , paneth cells
Carboxylated - sialoglycoprotein
Sialomucins - fetal mucin , upper part of crypts

Sulfated and Caboxylated - colonic mucins in goblet cells

PAS - POS
AB - POS

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5
Q

Group 4 carbs are

A

Glycolipids

Cerebrosides - lipid attached to carbohydrates

phosphatides - non carb containing lipids

PAS - POS

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6
Q

What are the stains that demostrate mucins

A

MMM
AB
PAS
Colloidal Iron

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7
Q

What is the purpose of alcian blue stainings

A

-demonstation of acid mucins (mucopoly) and sialomucins (glycoprotein)
-group 2 and 3 carbohydrates
-bind with ionic bonding or salt linkage

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8
Q

What are the control tissues for AB

A

Positive control - small intestine and appendix ANY GI

if ABPAS is used - duodenum is the best tissue because the brunners gland with neutral poly that pick up the PAS staining

always in include a small section of the kidney - most sensitive control for PAS

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9
Q

What are the characteristics of AB

A

-powder
-has copper phthalocyanin - basic dye with positive charge
-water soluble
-blue color because of the copper

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10
Q

how is the AB solution made

A

-powder dissolved in 3% acetic acid pH 2.5
-demonstrates sulfated and caboxylated acid mucopolys and sulfated/carboxy sialomucins

-if its made to pH 1.0 itll be ONLY sulfated acid muco and sulfated sialomucin

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11
Q

What are the advantages of Alcian blue over other cationic dyes

A

water, alcohol, weak acids and other counter stains will not extract the dye from stained sections
-the dye is strong
-incubaction time is adjustable
-dye can be combined with PAS/Hx to demonstrate neutral mucins AND acid mucins

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12
Q

What is ionic bonding

A

bond between cation and anion

staining depends on the charge of the dye and the charge of the tissue

-opposites attract

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13
Q

Acid dyes ANIONIC are _____ they bind to ___charged tissues and the tissue is described as

A

Acid dyes are NEGATIVE they bind to POSITIVE charged tissues and the tissue is described as ACIDOPHILIC

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14
Q

BASIC - CATIONIC dyes are _____ they bind to ___charged tissues and the tissue is described as

A

BASIC - CATIONIC dyes are POSITIVE they bind to NEGATIVELY charged tissues and the tissue is described as BASOPHILIC

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15
Q

What are amphoteric dyes

A

both basic and acidic groups behaving as both above or below a certain pH known as a isoelectric point

at pH 4 stains cytoplasm (positive charged) so it is an acid dye at pH 4 or below

protein tissue can also be amphoteric - 6-6.5pH
below the isoelectric point it is acidophilic or positively charged and above the pH its basophilic

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16
Q

How would protein staining on an acid solution work

A

if acid dye is negatively and is put on a protein in an acidic enviroment in low pH = BRIGHT Color because there are more reactions between the dye and tissue

as pH increases the intensity is weaker

If Basic dye is applied to protein in acid environment = weaker color and as pH goes up the intensity of the color goes up

17
Q

How would protein staining work in a basic solution

A

Basic dye is more intense in basic solution

Acid loses intensity as the solution gets more basic

18
Q

What happens if Alcian blue is added to tissues (cryts of lieberkuhn CL ) at different pH

A

1- little blue reaction
2-stronger
3-stronger
4-6 no staining in CL but other cells were stained
7 and up - no staining anywhere

AB does not stain mucipoly at pH of 4 or up

Acts as a basic dye at pH 1-4 , acid dye at pH 4-7 = amphoteric dye

19
Q

Why is there a 2nd 3% acetic acid rinse in AB staining

A

prevent non specific staining
dont want to change pH while stain is on the slide so under acetic acid before rinsing in tap water

20
Q

What is the step 1 of AB staining

A

acetic acid rinse
-same solvent as alcian blue solution
-protects AB staining solution from pH changes after being exposed to water

21
Q

Step 2 of AB staining

A

Staining
-done at pH 2.5 or 1.0
-amphoteric dye
-progressive stain and direct
-binds to negatively charged mucopoly and sialomucins which are caroxy/sulfated at pH 2.5 and sulphated at pH 1.0 (if pH 1.0 ordered then instead of acetic acid use HCL acid)
-if ABPAS because the neutral carbs have no charge they wont stained
-creates insoluble end product

22
Q

step 3 of AB staining

A

Counter staining

-ionic bonding
-demonstrates other tissue components
-has to be a contrasting color
-diff counter stains can be used
Hx, PAS, NFR, VG
-Most common is Nuclear Fast Red

23
Q

What would be a result of the staining

A

Expect: Mucin stained and counterstain should enhance the mucin

@ph 1.0
sulphated acid mucopoly and glycoproteins - pale blue
Background - pink to red

@pH 2.5
Weakly acidic mucopoly, hyaluronic acid and sialo - dark blue
background - pink to red

24
Q

pg 33 what ph

A

2.5
Nuceli are red

25
Q

34

A

ph 1

26
Q

35

A

ph 2.5

27
Q

37

A

neutral mucins vs acid mucin
2.5 (acid) first then PAS stain (neutral)
duodenum
brunners gland top left

Hem Yr 2 Sem 2 (6 decks)

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