Preparation of different Stains and Reagent Used for the diagnosis of fungal infections

M4S • Digital Textbook

Mycology • Laboratory Preparation

Preparation of Different Stains and Reagents Used for the Diagnosis of Fungal Infections

Advanced M4S Digital Textbook • Prepared by M4S

4 Preparations Mycology Laboratory Notes Interactive Study

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Contents

1. 10% KOH 2. 20% KOH 3. Giemsa Stain 4. Lactophenol Cotton Blue

1

10% a KOH

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Requirements

KOH pellets

Prodecure

• 10 gm of KOH pellets is weighed.
• Mixed with 100ml of distilled water.

2

20% KOH

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Requirements

KOH pellets

Procedure

• 20 gm of KOH pellets is weighed.
• Mixed with 100ml of distilled water.

3

Giemsa Stain

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Requirements

1. Giemsa Powder
2. Methanol
3. Glycerol

Procedure

• 3.8 gm Giemsa powder is transferred to 500 ml brown bottle containing few dry glass beads.
• 250 ml of Methanol and same volume of Glycerol is mixed well.
• The mixture is placed in water bath at 50-60 degree centigrade or at 37 degree centigrade for up to 2 hrs for complete dissolution.

4

Lactophenol Cotton Blue

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Requirements

1. Methyl blue (Cotton Blue)
2. Phenol Crystals
3. Lactic Acid
4. Glycerol

Procedure

• 0.04 gm of methyl blue is dissolved in 10 ml warm water.
• 10 gm of phenol crystals is added.
• 10 ml of lactic acid and 20 ml of glycerol is added and mixed properly.

M4S – Microbiology for Students
Preparation of Different Stains and Reagents Used for the Diagnosis of Fungal Infections