Method of staining
- First of all, labelled the slide and bring it to the water, it means put in to descending order of alcohol jars ( (95%,...
- Then stain with 1% Alcian blue solution for 15 – 30 minutes and after that rinse with distilled water.
- Then treat with safranin or neutral red for few seconds.
- Neutral red or safranine is used as counter stain.
How do you use Alcian blue?
For Alcian blue pH 1.0 or 0.2 use section of cartilage. Method. Bring sections to distilled water. Stain in the Alcian blue solution 15 mins. Wash well in running tap water 5 mins (For lower pH solutions ie pH 1.0/0.2, drain and blot dry, to prevent removal of stain in water). Rinse in distilled water. Counterstain with neutral red stain 1 min.
What is the composition of Alcian blue stain?
Alcian blue stain is mainly comprised with 1% Alcian blue, Glacial acetic acid and distilled water. For preparing of 100ml solution it should be used 0.5g of 1% Alcian blue dye powder, 3ml of glacial acetic acid and 97ml of distilled water.
What is the pH of Alcian blue solution?
3% Acetic Acid Solution: Alcian Blue Solution (pH 2.5): Mix well and adjust pH to 2.5 using acetic acid. Nuclear fast red ------------------- 0.1 g Dissolve a luminum sulfate in water. Add nuclear fast red and slowly heat to boil and cool.
Does oesophagus show Alcian blue stain?
Under normal conditions, a biopsy specimen of the oesophagus will have no staining on the alcian blue stain test. However, an intestinal biopsy specimen will have intestinal-type of goblet cells that contain acidic mucus, and hence will show positive staining with the alcian blue stain.
What is alcian blue made of?
PRINCIPLE: Alcian blue is a group of polyvalent basic dyes that are water soluble. The blue color is due to the presence of copper in the molecule. The 3% acetic acid solution (pH2. 5), Alcian blue stains both sulfated and carboxylated acid mucopolysaccharides and sulfated and carboxylated sialomucins (glycoproteins).
What is alcian blue technique?
Alcian blue may be used in combination with the PAS staining procedure, so that both acid and neutral mucins can be demonstrated in the same tissue sample. Alcian blue will stain acidic mucins blue and PAS will stain neutral mucins rose red. This technique is also useful in diagnosing diseases of the GI tract.
How do you use Alcian blue stain?
MethodDeparaffinize in xylene and rehydrate through graded ethanols to deionized water.Stain in the alcian blue solution for 30 minutes.Rinse in running tap water for 5 minutes.Counterstain in nuclear fast red for 10 minutes.Wash in running tap water for 1 minute.Dehydrate in graded ethanols.
What Colour does Alcian blue stain?
The Alcian blue at a pH of 2.5 will stain all acid mucins deep blue but will not color the neutral mucins. The subsequent application of the PAS technique will stain the neutral mucins bright magenta.
What does Alcian blue stain detect?
pH 2.5 alcian blue is used to diagnose Barrett's esophagus, as it will also stain goblet cells. It is also used in mesothelioma, atherosclerosis and adenocarcinomas.
What stain is used for mucin?
PAS (periodic acid-Schiff) It stains glycogen, mucin, mucoprotein, glycoprotein, as well as fungi.
Is alcian blue cationic or anionic?
Cationic dyeAlcian Blue 8GX, Cationic dye (ab145250)
What is mucin production?
Mucin glycoproteins produced by mucus-producing cells in the epithelium or submucosal glands are the major macromolecular constituent of mucus and are responsible for the viscous properties of the mucus gel.
Which tissue types are commonly stained with alcian blue?
1) Alcian BlueTissue typeCompositionAlcian blue 2.5Goblet cellsAcid mucins - SimplePositiveTissue StromaAcid mucins - SimplePositiveAdenocarcinomasAcid mucins - ComplexNegativeCartilage, BoneAcid mucins - ComplexNegative2 more rows
Why PAS staining is done?
The periodic acid-Schiff (PAS) staining procedure is most commonly used in the histology laboratory to detect glycogen deposits in the liver when glycogen storage disease is suspected. Glycogen granules may also be visible in tumors of the bladder, kidney, ovary, pancreas, and lung.
What is Van Gieson stain used for?
Van Gieson's trichrome is used for the differentiation between collagen and smooth muscle in tumors and to demonstrate the increase in the amount of collagen in these pathologies.
What is Mucicarmine stain used for?
Mucicarmine (Mucin Stain) is intended for use in the histological visualization of acid mucopolysaccharides in tissue sections. This product is useful in distinguishing mucin negative undifferentiated squamous cell lesions from mucin positive adenocarcinomas.
Introduction
Alcian blue is a special stain that is used to identify acid mucin and mucopolysaccharides.
Purpose
In histology alcian blue staining method is used to identify acid mucin and mucopolysaccharides seen in mesotheliomas, blood vessel walls and early lesions of atherosclerosis.
Principle
Alcian blue is basic dye containing positively charged groups comprised with copper molecules that gives it the blue colour which form salt linkage with poly anions (sulfated and carboxylated acid mucin and acid mucopolysaccharides).
Alcian blue solution
Alcian blue stain is mainly comprised with 1% Alcian blue, Glacial acetic acid and distilled water.
Method of staining
First of all, labelled the slide and bring it to the water, it means put in to descending order of alcohol jars ( (95%, 80%,70%, 60%, alcohol solution) and finally to the distilled water jar.
Uses
Alcian blue stain is really important for detection of acid mucin and acid mucopolysaccharides in connective tissues & epithelial tissues.
What is the color of the tissue that is stained with alcian blue?
The tissue parts that specifically stain by this dye become blue to bluish-green after staining and are called "Alcianophilic" (comparable to " eosinophilic " or " sudanophilic "). Alcian blue staining can be combined with H&E staining, PAS staining and van Gieson staining methods.
What is Alcian blue?
Alcian blue ( / ˈælʃən /) is any member of a family of polyvalent basic dyes, of which the Alcian blue 8G (also called Ingrain blue 1, and C.I. 74240, formerly called Alcian blue 8GX from the name of a batch of an ICI product) has been historically the most common and the most reliable member. It is used to stain acidic polysaccharides such as ...
What is the Monstral Blue?
The Monstral blue found to coat the inside of copper vessels used to process phthalic acid derivatives had led to the discovery of Phthalocyanine in 1907. Attracted by the brilliance, stability and insolubility of this chromophore, attempts were made to reversibly modify it so that it would be carried into fabric in a solution and then easily precipitated (ingrained) into an unleachable but finely well dispersed deposit (hence the name " ingrain dyeing "). From this attempt, Alcian blue (Ingrain blue 1) was first synthesized by the ICI dyestuffs department under N. H. Haddock and C. Wood in the early 1940s and patented in 1947, originally as a textile dye. In 1950 it was used by Steedman as a selective dye for mucins. While the popularity of Alcian blue expanded exponentially, the difficulty involved in its production due to environmentally hazardous intermediate steps made its availability difficult and ICI stopped producing it by 1973. Many of the alternate sources sold similar looking color products with unreliable staining.
What is the most widely used cationic dye for light and electron microscopy?
For many of these targets it is one of the most widely used cationic dyes for both light and electron microscopy. Use of alcian blue has historically been a popular staining method in histology especially for light microscopy in paraffin embedded sections and in semithin resin sections.
How long does Alcian Blue 8G last?
Acidic solutions of alcian blue 8G are often stable for some years. Churukian's lab manual [1] gives a recommended shelf life of 6 months. An alcian blue solution with a precipitate should be discarded and replaced, not filtered and used. Some dyes sold as alcian blue 8G are unstable in solutions at pH 5.6 and above; they precipitate in less than 24 hours. Batches of alcian blue that do not form stable solutions cannot be used in Scott's "critical electrolyte concentration" methods for histochemical characterization of different glycosaminoglycans, which require solutions at pH 5.7-5.8 with variable concentrations of MgCl 2 .
How many nm does Alcian blue have?
Thus even at a fairly high dilution, it has an absorption maximum at ~600–615 nm, which is actually not the absorption maximum of a dye monomer but that of the multimer.
When did ICI stop producing Alcian blue?
While the popularity of Alcian blue expanded exponentially, the difficulty involved in its production due to environmentally hazardous intermediate steps made its availability difficult and ICI stopped producing it by 1973. Many of the alternate sources sold similar looking color products with unreliable staining.
