Staining of Fungus by using Lactophenol cotton blue
Staining of Fungus by using Lactophenol cotton blue Aim To stain fungal hyphae by using Lactophenol cotton blue. Requirement -Young fungal culture -Glass slide -Cover slip -Needle -Lactophenol cotton blue stain. Procedure
1- Take a clean grease free slide. 2- Add a drop of mounting fluid that is lactophenol cotton blue solution on a slide. 3- Sterilize the needle and cool it then transfer a mycellial mat on fluid and press it gently so that it easily mix with the stain. 4- Take a clean cover slip and with the help of a forceps place the cover slip on mycellial mat. 5- Take a blotting paper and wipe the excess stain . 6- Observe under low to high power objectives of microscope. Principle: The slide culture is the best method for preserving and observing the actual
structure of a fungus. Materials: Actively growing fungus on Potato Dextrose agar. 1. Petri dish 2. Sterile water 3. Potato Dextrose agar 4 Coverslip 5. Glass slide 6. Lactophenol cotton blue Procedure: 1. Place a slide on a bent-glass rod and a piece of filter paper in the bottom of a petri dish
2. Prepare a drop of potato dextrose agar by using sterilized pipette in the center of the slide in a petri dish and allow to solidify. 3. Inoculate the sides of PDA agar drop with a small fragment of fungus being studied. 5. Sterilize coverslip by passing it briefly through a flame. Cover inoculated drop with the sterile cover slip. 6. Add sterile water to bottom of petri dish. 7. Incubate at 25 C until sporulation occurs (usually 1-2 weeks, occasionally faster). The slide preparation may be checked periodically under the low power of a microscope. 8. When sporulation is complete, carefully lift off cover slip and place onto a drop of lactophenol cotton blue on a clean slide, making your first slide.
NOTES -Phenol will kill any live organisms -Lactic acid preserves fungal structures -Lactophenol cotton blue stains the chitin in the fungal cell walls. Making a wet mount slide and observing yeast under microscope Aim To prepare a wet mount of yeast cell and to observe them under the microscope
Materials -Yeast cell suspension -Glass slides -Cover slip -Needle -safranin Procedure 1- Take a sample of yeast suspension provided and add a single drop to a clean microscope slide 2- Place a cover slip on the drop without causing air bubbles 3- Observe the yeast cells under low magnification at first and under high power
4- Stain the cells with safranin Results Draw a picture of the cells that you see on the wet mount slide. Saccharomyces Schizosachromyces Rhodotorula Hansenulla
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