India ink capsule staining is an effective method for observing Cryptococcus neoformans. India ink is a high-quality reagent for this assay. Cryptococcus neoformans is surrounded by a mucopolysaccharide capsule, which can be clearly demonstrated via India ink staining. After staining, the capsule excludes colloidal carbon particles in the ink, and the fungal cells are surrounded by distinct, colorless and transparent halos.
Instructions for Use:
1. Centrifuge the sample, collect the pellet and spread it evenly on a glass slide, then add a drop of India ink onto the smear.
2. Place a coverslip carefully to avoid air bubbles. Gently press the coverslip with your finger to spread the sample and ink into a thin layer.
3. Screen for encapsulated microorganisms under a low-power objective, then observe and identify in detail under a high-power objective or oil immersion objective.
Precautions:
1. Use opened India ink within 6 months. Seal the container tightly after each use to prevent volatilization and deterioration.
2. Precipitate may form after long-term storage of India ink. If coarse particles are observed under the microscope, filter the ink before use or discard it.
3. Ensure appropriate purity and concentration of the sample. A too-low concentration will make Cryptococcus neoformans difficult to detect microscopically.
4. Distribute the sample and India ink uniformly on the slide.
5. Dilute the ink slightly if it is overly concentrated.
6. Wear a lab coat and disposable gloves during operation for personal safety and health protection.
Staining Results:
Cryptococcus neoformans.
| Surrounded by a thick, transparent capsule with clearly visible cell walls
|
Background
| Black |
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