Negative control: Absence of encapsulated yeastsĬapsules of Cryptococcus neoformans like organisms seen Importance of India Ink Preparation Assay.Positive control: Presence of encapsulated yeasts.Result Interpretation of India Ink Preparation The encapsulated strain of Cryptococcus neoformans in India Ink preparation of CSF The encapsulated strain of Cryptococcus neoformans in India Ink preparation of CSF Look for oval or round cells, some showing budding, irregular in size, measuring 2-10 µm in diameter, and surrounded by a large unstained capsule as shown in the figure. If any suspicious objects encounter, examine with the high dry objective (40X).Examine the entire 22- by 22-mm coverslip systematically with the low power objective (10X ) and low light intensity.a drop of the sediment to a slide and add a drop of India ink. Transfer an equal amount of sediment and India ink i.e.Remove the supernatant fluid and mix the sediment.Centrifuge the CSF for 5 to 10 minutes.Control stains (For positive control- Cryptococcus neoformansĪnd for negative control Candida albicans).Clean and grease-free slide and coverslips.
Therefore, capsules appear as a clear halo around the yeast cells. The capsules are non-ionic so the India ink used will not bind to it. India Ink Preparation Step Principle of India Ink India ink is used as a negative stain in negative staining that uses ion negative staining technique permits visualization of the usually transparent and unstainable capsules of various micro- microorganisms like Cryptococcus neoformans (most commonly), Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae, etc. The capsulated strain of Haemophilus influenzae in Gram-staining of sputum showing pleomorphic Gram-negative coccobacilli to small and large rods and bodies surrounded by clear zones at a magnification of 2000XĬryptococcal meningitis occurs in immunodeficient patients and when meningitis is clinically suspected, for example, patients with HIV/AIDS, or when yeast cells with lymphocytes are detected when performing Cerebrospinal fluid (CSF) cell count or examining Gram smear, examine India ink preparation for encapsulated yeasts.Capsulated strain of pneumococcus in Gram staining of sputum showing Gram-positive diplococci and bodies surrounded by clear zones.Encapsulated Streptococcus pneumoniae in Gram staining of CSF showing Gram-positive diplococci bodies with clear zones.Capsule of Klebsiella pneumoniae in negative staining or dry India ink preparation microscopy.Capsules and yeast cells of Cryptococcus neoformans in negative staining of culture microscopy at a high magnification of 1600X.Capsules and yeast cells of Cryptococcus neoformans in negative staining of CSF microscopy at a high magnification.Capsules of Cryptococcus neoformans in Nigrosin wet mount microscopy.Cryptococcus Capsules in Negative Staining or Nigrosin Preparation.Limitations of Ink Ink or Negative staining.Importance of India Ink Preparation Assay.The encapsulated strain of Cryptococcus neoformans in India Ink preparation of CSF.
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