Lactophenol Cotton Blue stain principle and interpretation for Fungi specimen

LACTOPHENOL COTTON BLUE MTGEEKS

Fungi are members of  the eukaryotic organism and they are classified into two main groups that is yeast and molds.Those fungi capable of exhibiting this to stages are known as dimorphic fungi. Fungi cell wall is made up of chitin and the cells have both macroscopic as well as microscopic structure.

Lactophenol Cotton Blue reagent is used for fungal staining.

The lactophenol cotton blue abbreviated lpcb stain wet mount preparation is one of  the most widely used method of staining and observing fungi because it is simple to prepare.

The identification of molds is based on the shape, method of production, and arrangement of spores (conidial ontogeny). lactophenol blue stain Solution is a mounting medium and staining agent used in the preparation of slides for microscopic examination of fungi.It intensely stain fungi elements blue.

Principle of Lactophenol Cotton Blue stain

The preparation of Lactophenol cotton blue stain  has three components thus

  • Lactic acid preserves the cellular  structure of fungi and clears the tissue
  • Phenol acts as a disinfectant and cotton blue imparts blue colouration to the fungal spores and hyphae

Composition of Lactophenol Cotton Blue stain

The following ingredients are used for the composition Lactophenol cotton blue stain

IngredientsIn 100 ml aqueous solution
Phenol crystals20.000
Cotton blue 0.050
Lactic acid20.000
Glycerol20.000
Distilled water20.000

How to Prepare Lactophenol cotton blue

  • Start by placing a drop of Lactophenol Cotton Blue reagent on a clean and dry slide.
  • The stain imparts a blue colouration on hyphae.
  • By using a nichrome inoculating wire, carefully spread the fungal culture into a thin preparation
  • Now place a coverslip on the thin preparation.
  • Then place a coverslip edge on the drop and slowly lower it. Avoid trapping air bubbles under the coverslip and allow  the stain to act for about 5 minutes.
  • Observe under microscope with low power 40X objectives  lens for screening in low intensity.
Lactophenol slide
Lactophenol slide
Lactophenol slide covered
Lactophenol slide covered

A considerable discoloration of the original  fungi material can be obtained by treating it with KOH before staining.

Also read  Albert staining of metachromatic granules ideal for Corynebacterium diphtheriae

Appearance of Lactophenol cotton blue stain

It has an Ink blue coloured solution

lactophenol cotton blue stain
lactophenol cotton blue stain

Type of Inoculums

Primarily with pure cultures that’s is clinical samples  such as

Observation and interpretation of Lactophenol cotton blue stain

Fungal spores and hyphaepale to dark blue

Quality Control Organism

Aspergillus nigerDelicate blue hyphae and fruiting structures with a pale blue background.
Trichophyton mentagrophytesDelicate blue hyphae and fruiting structures with a pale blue background.
aspergillus species in lactophenol stain
aspergillus species in lactophenol stain
Lactophenol Cotton Blue Stain of Penicillium Species

Lactophenol Cotton Blue Stain of Penicillium Species
Lactophenol cotton blue showing Chrysosporium species
Lactophenol cotton blue showing Chrysosporium species
Lactophenol cotton blue mount of the isolate showing  (a) oval to ellipsoidal aleuriconidia, (b) arthroconidia and (c) chlamydospores.
Lactophenol cotton blue mount of the isolate showing (a) oval to ellipsoidal aleuriconidia, (b) arthroconidia and (c) chlamydospores.

Possible limitation of Lactophenol cotton blue stain

The stain is useful in the recognition and presumptive identification of fungi.It is advice to do some biochemical test in order to observe or identify additional characteristics including colony morphology where appropriate for final identification and conclusion

Reference

  • Generalmicroscience.com/microbiallaboratorytechniques/staining-fungus-using-lactophenol-cotton-blue.
  • Koneman, E.W., et al. Color Atlas and Textbook of Diagnostic Microbiology, J.B. Lippincott Company, Philadelphia, PA.
  • Isenberg, H.D. Clinical Microbiology Procedures Handbook. 2nd Edition.
  • Jorgensen,J.H., Pfaller , M.A., Carroll, K.C., Funke, G., Landry, M.L., Richter, S.S and Warnock., D.W. (2015) Manual of Clinical Microbiology, 11th Edition. Vol. 1.

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About the Author: Arthur Westmann

DEFFE ARTHUR (AMOEBAMANN) is the founder and author of MLTGEEKS and MLTEXPO.He’s from Cameroon and is currently a Final year State Medical Laboratory Technician (MLT MA). Beyond lab works, he’s a passionate internet user with a keen interest in web design and blogging. Furthermore He likes traveling, hanging around with friends and social networking to do in his spare time.

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