1. Slide culture
Dalia Kamal Eldien
Practical NO(4)

2. Slide culture
Special techniques used in mycological lab include Slide culture and Scotch tape mount
In slide cultures, we are growing the fungi directly on the slide on a thin film of agar.
By doing this, there is no need to remove a portion of the fungus from a culture plate and transfer it to the slide.
So there is  less chance for the fungi features that are key to identification, notably the spore-bearing structures, to be damaged.

3. Method
Aseptically, with a pair of forceps, place a sheet of sterile filter paper in a Petri dish.
Place a sterile U-shaped glass rod on the filter paper.
Pour enough sterile water  on  filter paper to completely moisten it.
With forceps, place a sterile slide over the U-shaped rod
Gently flame a scalpel to sterilize, and cut a 5 mm square block  of  the medium  from  the  plate  of Sabouraud’s agar .

4. Method
Pick up the block of agar by inserting the scalpel and carefully transfer this block aseptically to the centre of the slide.
Inoculate four sides  of the agar square with spores or mycelial fragments of the fungus to be examined.
Aseptically, place a sterile cover glass on the upper surface of the agar cube.

5. Method
Place the cover on the Petri dish and incubate at room temperature for 48 hours.
After  48  hours,  examine  the  slide  under  low power. If growth has occurred there will be growth of hyphae  and production of  spores.

6. Application of Stain
Place a drop of lactophenol cotton blue stain on a clean microscope slide.
Remove the cover glass from the slide culture and discard the block of agar.
Add a drop of 95% ethanol to the hyphae on the cover glass. As soon as most of the alcohol has evaporated  place  the  cover  glass,  mold  side down,  on  the  drop  of  lactophenol  cotton  blue stain on the slide.  Examine the slide under microscope

7. Cut the block of agar

8. Pick up the block of agar

9. U-shaped glass rod

10. Wet the filter paper

11. Culture the tested fungi

12. Cover by cover slip

13. Growth of fungi after incubation

14. Examine under microscope

16. Scotch tape mount
The Scotch tape mount is an easy and fast procedure that is used for the identification of filamentous fungi since most structures will be intact for observation thank to the gummed side of the tape
Cut a strip of Scotch transparent tape and place ends between thumb and index finger, gummed side out.
Open plate with opposite hand and press tape against the colony to identify.

17. Place a drop of lactophenol on a labeled slide, Press tape against slide with lactophenol.
Smooth the tape back on the slide by opening fingers and using guaze.
Place another drop of lactophenol on top of the tape.
Place a large 20x40 mm coverslip on top of slide.
Examine the slide under the microscope.

18. Scotch tape

20. Scotch tape preparation

21. Examine under the microscope