(LACTOPHENOL BLUE STAINING)
: 1) FARAH ANINI BINTI MD. SHAFI’I
2) FATIN NUR SYAHIRAH BINTI MOHD. HANAFI
3) NOR AZAH BINTI MAHIZAN
4) JOSHUA RAJ A/L ISIDORE SAKAYARAJ
5) CHAN POH SHIN
: 1) 012012051710
: BACHELOR OF NUTRITION (BNT)
: MADAM SASIMALANI SARGUNNAM
Figure 1: Common fungi
The Kingdom Fungi includes some of the most important organisms, both in terms of their ecological and economic roles. By breaking down dead organic material, they continue the cycle of nutrients through ecosystems. In addition, most vascular plants could not grow without the symbiotic fungi, or mycorrhizae, that inhabit their roots and supply essential nutrients. Other fungi provide numerous drugs such as penicillin and other antibiotics, foods like mushrooms, truffles and morels, and the bubbles in bread, champagne, and beer. Fungi also cause a number of plant and animal diseases: in humans, ringworm, athlete's foot, and several more serious diseases are caused by fungi. Fungi are more chemically and genetically similar to animals than other organisms, thus this makes fungal diseases very difficult to treat. Plant diseases caused by fungi include rusts, smuts, and leaf, root, and stem rots, and may cause severe damage to crops. However, a number of fungi, in particular the yeasts, are important "model organisms" for studying problems in genetics and molecular biology. The lactophenol blue staining or known as lacto phenol cotton blue (LPCB) wet mount preparation is the most widely used method of staining and observing fungi and is simple to prepare. The preparation has three components: phenol, which will kill any live organisms. The high concentration of the phenol deactivates lytic cellular enzymes thus the cells do not lyse. Meanwhile, lactic acid which preserves fungal the structures and cotton blue which stains the chitin in the fungal cell walls. Lactophenol cotton blue is recommended for mounting and staining yeast and molds. By staining fungi with lacto phenol a microbiologist can clearly see it under the microscope. The cotton blue lactophenol is the most common method of staining fungi. The dye will penetrate the hyphae and reproductive cells in the fungi.
PRINCIPLE OF LACTOPHENOL COTTON BLUE STAINING
The Lactophenol Cotton Blue preparation is used to stain fungal elements, which aids in the microscopic identification of mycotic agents. Lactophenol Cotton Blue Stain is formulated with lactophenol, which serves as a mounting fluid, and cotton blue. Lactophenol Cotton Blue is recommended for mounting and staining yeast and moulds. Organisms suspended in the stain are killed due to the presence of phenol. The high concentration of the phenol deactivates lytic cellular enzymes thus the cells do not lyse. Cotton blue is an acid dye that stains the chitin a nitrogenous substance present in the cell walls of fungi. Lactophenol Cotton Blue Staining is intended use for mounting and clearing skin scrapings or hair shafts and to aid in the microscopic visualization of fungal elements in these specimens. This staining can also be used to examine fungal colonies microscopically for morphology, or permanent mounting of fungal specimens.
Lactic Acid has long been used as a clearing agent for plant tissue. The combined lactic acid, phenol and aniline (cotton) blue was used to visualize the cellular structure in plant material. It was later found that the combined mixture which is the Lacto-phenol cotton blue stained fungal elements very well too. Clearing, staining and mounting is achieved with skin scrapings and hair shafts in one step. The sample for staining can be taken from skin or hair and fungal cultures (small part of colony growing on suitable media).
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