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IBEX SCIENCES is a well-established laboratory chemical, glassware, and scientific apparatus supplier based in Lahore, Pakistan. They provide critical laboratory reagents, including Leishman Stain Solution, for clinical diagnostics, research laboratories, and educational institutions. [1, 2]
Overview & Diagnostic Purpose
Leishman Stain Solution is a specialized Romanowsky-type differential histological stain. It is primarily formulated for the standard microscopic examination of blood smears and bone marrow samples to evaluate cellular morphology. In clinical hematology and pathology, it serves as a frontline tool for: [1, 2, 3, 4]
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- Leukocyte Differentiation: Distinctly coloring white blood cells (neutrophils, lymphocytes, eosinophils, basophils, and monocytes) to perform differential cell counts. [1, 2, 3]
- Parasite Detection: Identifying blood-borne protozoa, specifically Plasmodium species (the causative agent of malaria) and trypanosomes. [1, 2, 3, 4, 5]
- Microbiological Screening: Assisting in the preliminary identification of specific bacterial structures in systemic infections. [1]
Chemical Composition & Core Principle
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- The Mixture: It contains a methanolic combination of polychromed methylene blue (a basic/cationic dye) and eosin (an acidic/anionic dye). [1, 2]
- Staining Affinity:
- Methanol Integration: The solution utilizes high-purity methanol as its solvent matrix. This provides a dual-action mechanism where the solution acts as its own cellular fixative. When applied to an un-fixed, dry blood film, the methanol stabilizes cell morphology instantly, eliminating the requirement for a separate formal fixation stage. [1, 2]
Standard Laboratory Procedure
To achieve highly reproducible and artifact-free diagnostic results, the stain is typically applied following a strict timing sequence:
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- Smear Preparation: Spread a thin, clean blood film across a high-quality glass slide using the wedge technique and let it air-dry completely. [1]
- Fixation: Flood the un-fixed slide with undiluted Leishman Stain Solution for 1 to 2 minutes. This step fixes the blood cells onto the glass slide. [1, 2]
- Dilution & Staining: Add double the volume of pH-buffered distilled water (ideal pH 6.8 to 7.2) directly to the slide. Mix gently until a faint metallic scum forms on the liquid surface, and leave to incubate for 10 to 12 minutes to allow complex dye interaction. [1, 2]
- Rinse & Dry: Wash the slide briefly with buffered or running distilled water until the film takes on a bright pinkish translucent appearance. Drain and allow to dry in a vertical position. [1]
- Microscopy: Examine under an optical microscope using a 100x oil-immersion objective lens for full evaluation. [1]
Storage and Lab Handling Guidelines
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- Evaporation Management: The bottle must be stored tightly sealed. Methanol is highly volatile; evaporation can skew the dye ratio and lead to dense precipitation, creating heavy background debris on your diagnostic slides.
- Flammability Risk: Because of the high methanol content, the solution is highly flammable and must be kept clear of heat sources, sparks, or open lab flames.
- Health Precautions: The solution is toxic if inhaled, ingested, or absorbed through the skin. Always handle the liquid within a well-ventilated space or under a fume hood while wearing appropriate personal protective equipment (PPE). [1]
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