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.
Reference strains | Conditions | Expected results |
S. aureus ATCC 25923 | oil immersion lens (100x) | dark violet with gentian |
E. coli ATCC 35218 | pink-red with fuchsin |
- BacterStain™ Crystal Violet (Code: 04007)
- BacterStain™ Lugol (Code: 04005)
- BacterStain™ Safranine (Code: 04008)
- BacterStain™ Alcohol (Code: 04011)
*Note: This ingredient is for reference only our company will formulate the ingredient according to requests or tender specifications.
Product Name | Code | Packaging |
BacterStain™ Grams Stain Kit | 04001 | 4 bottles of 100ml/box or according to the customer’s request.. |
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