GBL® ROSA-STAIN

5060-Modified Alcyan Blue pH: 2.5 Staining Kit

Title Description
REF No 5060
Purpose It is used to examine acid mucins, carboxylated and sulfated mucous substances in tissue sections by light microscopy.
Ingredients A) Alcian Blue Stain pH: 2.5: Alcian blue, Glacial acetic acid
B) Nuclear Fast Red Stain: Nuclear fast red
C) Acetic Acid; 3%: Glacial acetic acid
Working Principle Alcian Blue dye interacts in a unique way with acidic mucopolysaccharides. These mucopolysaccharides are found mainly in mucous membranes and some other tissues. After interacting with acidic mucopolysaccharides, the dye forms a complex with these compounds and this complex is blue. This blue color, which is concentrated in mucosal areas, allows the mucosa to be highlighted.
Alcyan blue is a water-soluble polyvalent basic dye. The blue staining comes from the copper in the molecule. At the acid pH formed by acetic acid,
a)sulfated acid mucopolysaccharides
b)carboxylated acid mucopolysaccharides
c)sulfated sialomucins (glycoproteins)
d) Carboxylated sialomucins (glycoproteins) stain with alcyan blue.
Warnings, Precautions, Recommendations Danger Caution. Causes serious eye damage. Causes skin irritation. Wear protective gloves/protective clothing/eye protection/face protection. IN CASE OF CONTACT WITH EYES: Rinse carefully with water for several minutes. Remove contact lenses if fitted and easy to do so. Continue rinsing. IN CASE OF CONTACT WITH SKIN: Wash with plenty of soap and water. In case of skin irritation: Get medical advice/attention.
Application SAMPLES AND PREPARATION
1. Fixation: Especially with formalin fixatives; Bouin and Hollande fixatives can also be used.
2. Staining Method: Overlay or immersion.
3. Control: Small intestine, appendix and colon.
4. Warning-1: If mercury fixatives are used, mercury should be removed using iodine and thiosulfate solutions before staining.
5. Warning-2: The tissue section on the slide should be rinsed in distilled water before staining. If picric acid fixatives were used, it should be washed with distilled water for a long time.
6. Deparaffinize and rehydrate sections in the conventional way. Fixated, paraffin-embedded tissue sections (3 - 5 µm thick paraffin sections) are used.
APPLICATION OF THE STAINING METHOD:
1. Pre-treatment: Soak the preparation in “C- Acetic Acid; 3%” for 3 minutes.
2. Wash gently with distilled water for 1 minute.
3. Cover the tissue section with 5-10 drops of "A- Alcian Blue Stain pH: 2.5". After 30 minutes, remove the reagent by gently shaking the slide.
4. Wash gently with distilled water for 3 minutes.
5. Cover with 5-10 drops of “B- Nuclear Fast Red Stain”. After 10 minutes, shake the slide to remove the reagent.
6. Wash gently with distilled water for 3 minutes.
7. Dehydrate, clean, cover with xylene-based cover medium and examine under a microscope.
Theoretical Results
Acidic mucins, carboxylated and sulfated mucosubstances Blue
Nucleus Red/Pink
Cytoplasm, background Pale pink

Related Media
Shelf Life 2 years
Shelf Life After Opening the Lid
Storage Temperature 15-25 °C
pH (at 25°C) Component A pH: 2,5
Transportation Information
Ordering and Packaging Information
Order No Unit Volume Packaging Type Quantity in a Box
445060050050 Piece 50 mL 50 ml, White Bottle with HDPE Dropper 1 kit

Keywords Acidic mucins, carboxylated and sulfated mucosubstances, acid mucin staining, carboxylated mucin staining and sulfated mucous staining
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