Aspergillus flavus: LPCB Preparation and Observation of Its Structures
Aspergillus flavus in LPCB Mount
Aspergillus flavus and its structures in lactophenol cotton blue (LPCB) mount as shown above image and LPCB uses for staining and microscopic identification of fungi.
Principle of LPCB Mount Preparation
LPCB is used both as a mounting fluid and a stain. Lactic acid acts as a
clearing agent and aids in preserving the fungal structures; phenol kills the
organism and fixes it; glycerol prevents drying and cotton blue provides
color to the structure. It can be used alone or in conjunction with KOH.
Composition of LPCB
Lactic acid: 20 ml
Phenol: 20 ml
Glycerol: 40 ml
Cotton blue or aniline blue: 0.05 g
Distilled water: 20 ml
- Dissolve phenol in lactic acid, glycerol, and distilled water.
- After that, add cotton blue and mix well.
LPCB mount preparation procedure
- Take a clean grease-free glass slide.
- Put a large drop of LPCB with a Pasteur pipette.
- Transfer a small quantity of the culture to the drop.
- Tease the culture (in case of a mold) well with teasing needles so as to get a uniform spread.
- Put on a coverslip gently to avoid entrapment of air bubbles.
- Examine under low- (10X) and high-power (40X) objectives.
- Observe the morphological features like hyphae, conidiophore, vesicle, phialides, conidia carefully.
Further Readings
- Medical Mycology. Editors: Emmons and Binford, 2nd ed 1970, Publisher Lea and Febiger, Philadelphia.
- Rippon’s JW: Medical Microbiology. The pathogenic fungi and the Pathogenic Actinomycetes. 3rd ed 1988 Publisher WB Saunder co, Philadelphia.
- Clinical Microbiology Procedure Handbook Vol. I & II, Chief in editor H.D. Isenberg, Albert Einstein College of Medicine, New York, Publisher ASM (American Society for Microbiology), Washington DC.
- A Textbook of Medical Mycology. Editor: Jagdish Chander. Publication Mehata, India.
- Practical Laboratory Mycology. Editors: Koneman E.W. and G.D. Roberts, 3rd ed 1985, Publisher Williams and Wilkins, Baltimore.
- Topley & Wilsons Medical Mycology. Editors: M.T. Parker & L.H. Collier, 8th ed 1990, Publisher Edward Arnold publication, London.
- Textbook of Diagnostic Microbiology. Editors: Connie R. Mahon, Donald G. Lehman & George Manuselis, 3rd edition2007, Publisher Elsevier.
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