BacterStain™ Gram Stain Kit

A set of reagents used in a technique for the differentiation of Gram-negative and Gram-positive microorganisms.
Code: 04001
Packaging: Crystal Violet; Lugol; Safranin; Alcohol: 4 bottles/box 100ml or as per customer request.
Storage Conditions: 18–25°C.
Transportation: Ambient temperature.
Shelf Life: 24 months from the date of manufacture.

INTENDED USE

BacterStrain™ Grams Stain Kit is used for differentiation of bacteria on the basis of their gram nature.

PRINCIPLES

The gram staining kit consists of 4 bottles containing Crystal Violet, Safranine, Lugol (Iodine) and Alcohol (Decolourizer).
Crystal Violet, a basic dye, is the primary stain taken up by all bacteria due to its ability to rapidly permeate the cell wall. It stains the protoplast of bacteria purple. The potassium-iodine mixture (Lugol), in gram iodine, is the mordant which complexes with the primary stain in the cell. In gram-positive cells, the crystal violet-iodine complex is trapped in the cell due to a decrease in cell wall permeability caused by alcohol dehydration. Gram-positive cells are stained purple.
In gram-negative cells, the complex is removed by the Decolorizer due to an increase in permeability caused by solubility of the lipids in alcohol.
The counterstain used must be a contrasting color to the primary stain and in this case it is Safranin, which stains gram-negative cells red.

Prepare a thin smear on clear, dry glass slide.
Allow it to air dry and fix by gentle heat
Flood with BacterStain™ Crystal Violet (Code: 04007) for 1 minute.
Wash with tap water
Flood the smear with BacterStain™ Lugol (Code: 04005). Allow it to remain for 1 minute
Decolourize with BacterStain™ Alcohol (Code: 04011) until the blue dye no longer flows from the smear
Wash with tap water
Counter stain with BacterStain™ Safranine (Code: 04008) for 1 minutes and rinses off with water
Allow the slide to air dry or blot dry between sheets of clean bibulous paper and examine under oil immersion objective

RESULTS

Gram-positive bacteria stain dark violet with gentian: Staphylococcus, Streptococcus, Pneumococcus, Diphtheria bacillus, Bacillus anthracis.
Gram-negative bacteria do not stain violet with gentian, so they appear pink-red with fuchsin: Neisseria gonorrhoeae, Neisseria meningitidis, Escherichia coli, Shigella, Salmonella, Vibrio cholerae.

BacterLab ensures the quality of each product batch by testing with ATCC reference strains.

*Note: This ingredient is for reference only our company will formulate the ingredient according to requests or tender specifications.