Staining solution reagent kit IC400

for microbiologybacteria

Short description
Gram Staining Kit – 4 ready-to-use solutions × 250 ml – IVD – complete kit (crystal violet, Lugol, decolorizer, safranin) for differential bacterial staining. Gram-positive bacteria blue-violet; Gram-negative bacteria red-orange. Supplied by Pro-Eko Srl.

Description
The Gram Staining Kit is a diagnostic bacteriology kit composed of four ready-to-use solutions (250 ml each), supplied by Pro-Eko Srl as an in vitro diagnostic device (IVD). It allows complete execution of the Gram stain, the primary differential staining method in clinical microbiology, which classifies bacteria as Gram-positive (thick peptidoglycan layer, retains crystal violet–iodine complex) or Gram-negative (thin peptidoglycan layer with outer membrane, loses the complex during decolorization). The kit includes Lugol solution as mordant and is compliant with Ph. Eur. and referenced by ISS.

Kit composition

• Solution 1 — Gram Crystal Violet (0.5–1%): primary stain — stains all bacteria blue‑violet
• Solution 2 — Gram Lugol (iodine–iodide solution): mordant — forms the crystal violet–iodine complex
• Solution 3 — Decolorizer (acetone/ethanol): decolorization — removes the complex from Gram‑negative cells
• Solution 4 — Safranin O (0.25–0.5%): counterstain — stains Gram‑negative bacteria red‑orange
• Volume per solution: 250 ml × 4 (kit 4×250 ml)
• Product type: In vitro diagnostic medical device (IVD) — Reg. (EU) 2017/746

Gram staining principle and clinical importance
Gram staining exploits differences in bacterial cell wall structure: Gram‑positive organisms have a thick peptidoglycan layer that traps the crystal violet–iodine complex during alcohol/acetone decolorization, appearing blue‑violet; Gram‑negative organisms have a thin peptidoglycan layer and an outer membrane that permits loss of the complex, appearing red‑orange after safranin counterstain. Gram stain is recommended by Ph. Eur. and used as an initial, rapid test in clinical microbiology to guide empirical therapy while awaiting identification and susceptibility results.

Gram staining procedure

• Slide preparation: spread the sample (culture or clinical material) on a slide and fix by flaming or methanol
• Crystal violet: cover with 10 drops of Solution 1 — leave for 3 minutes
• Lugol: drain without washing — apply 10 drops of Solution 2 — leave for 1 minute (formation of the CV–I2 complex)
• Decolorization: drain — apply Solution 3 with agitation for 10–20 seconds until no more dye is released — immediately rinse with distilled water to stop decolorization
• Counterstaining: partially dry — apply 10 drops of Solution 4 — leave for 30–60 seconds
• Final wash: rinse with distilled water and dry with filter paper or air
• Observation: examine at 100× with immersion oil

Results
Gram‑positive = blue‑violet; Gram‑negative = red‑orange.

Application fields

• Clinical microbiology: rapid bacterial classification in infections (pneumonia, meningitis, sepsis, UTI, wound infections)
• Pharmaceutical quality control: microbiological testing of sterile and non‑sterile preparations (Ph. Eur.)
• Research laboratories: strain characterization
• Education: practical microbiology training
• Food industry: routine microbiological control of food samples

CLP classification and safety
The kit contains components with different CLP classifications: the decolorizer (acetone/ethanol) is flammable (H225); crystal violet solution may present H302; safranin has no harmonized classification. Use away from open flames during decolorization and consult the safety data sheets (SDS) for each component. IVD device pursuant to Reg. (EU) 2017/746.

Customisation and manufacturing
Pro-Eko Srl manufactures the Gram kit in alternative formats on request (for example 4×100 ml and 4×500 ml). Dedicated batches with IVD technical documentation are available for accredited facilities and quality control laboratories.

Technical specifications

• Manufacturer: Pro-Eko Srl
• Commercial name (model): Gram Staining Kit — SKU: IC400
• Composition: 4 ready‑to‑use solutions (Crystal Violet 0.5–1%; Lugol; Decolorizer acetone/ethanol; Safranin O 0.25–0.5%)
• Volume per solution: 250 ml × 4 (kit 4×250 ml)
• Estimated coverage: ~500–800 slides per 250 ml solution using 10‑drop method (approx.)
• Product type / Regulation: IVD — Reg. (EU) 2017/746
• Standards / Compliance: Compliant with European Pharmacopoeia (Ph. Eur.); referenced by ISS for clinical microbiology
• Weight (pack): 3 kg