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KOH Wet Mount Preparation: Purpose, Principle, Procedure, Result Interpretation and uses

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  Purpose:   To identify fungi in clinical specimens such as hair, skin or nails. Principle:   The KOH mount is used to aid in detecting fungal elements in thick mucoid material or in specimens containing keratinous materials such as skin scales, nails and hair.  The KOH dissolves the background keratin, unmasking the fungus element to make them more apparent. This test cannot find out the exact species of fungi. It can only differentiate whether fungi is present or not and the basic fungal structure. Further tests must be performed to confirm the species of fungi. Type of sample: Sterile containers should be used for sample collection Skin Hair Nail Requirements: Equipment Light microscope (40X magnification) Biological safety cabinet Reagents 10% KOH (aqueous) 40% KOH (aqueous) Cover slip  Microscopic slides Procedure for 10% KOH Preparation: Measure the 10g of potassium hydroxide (KOH) pellets, the transfer to container. Add 50ml distilled wa...
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India Ink Preparation- Purpose, Principle, Procedure and Result

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  Purpose:   Used for direct microscopic examination of capsules of many microorganism. Principle:   India ink or Nigrosin preparation is used for direct microscopic examination of capsules of many microorganisms .   The fine granules of the India ink or Nigrosin give an opaque background against which the clear capsules can be easily seen.  This technique is particularly useful in visualizing the capsule of  Cryptococcus neoformans  in cerebrospinal fluid, sputum and other secretions. Required equipment and reagents: Equipments: 1.    Bunsen burner 2.    Biological safety cabinet 3.    Light Microscope – 40 x magnifications Reagents: 1.    Cover slip 2.    Microscopic slides 3.    Nigrosin stain Procedure: 1.     Centrifuge CSF specimen lightly to concentrate any microorganisms in the specimen. 2.     Em...
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Oxidative-Fermentative Test (Hugh and Leifson)- Principle, Test Procedure Results and Interpretation

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  Objective To detect the oxidation or fermentation of carbohydrates by bacteria. Principle Saccharolytic microorganisms degrade glucose either fermentatively or oxidatively. The end products of fermentation are relatively strong mixed acids that can be detected in a conventional fermentation test medium. However, the acids formed in oxidative degradation of glucose are extremely weak, and the more sensitive oxidation-fermentation medium of Hugh and Leifson (OF medium) is required for their detection. The OF medium of Hugh and Leifson differs from carbohydrate fermentation media as follows: The concentration of peptone is decreased from 1% to 0.2%. The concentration of carbohydrate is increased from 0.5% to 1.0%. The concentration of agar is decreased from 1.5% to 0.3%, making it semisolid. The lower protein/carbohydrate ratio reduces the formation of alkaline amines that can neutralize the small quantities of weak acids that may form from oxidative metabolism. ...
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