Topic: The Fundamentals of Microscopy and technique of specimen preparation.
Objectives: To become familiar with the components of microscopes and become familiar with different methods to prepare the slide mounts for observation under the microscopes.
Materials:
1. Cultures of fungi with spores. Thielaviopsis basicola, C ladosporium sp., Alternaria brassicicola, Aspergillus niger and pythium ultimum
2. Olympus compound microscope. One is available with an attached camera.
3. A kit containing microscope slides, cover slips, needles, and transfer loops.
4. Immersion oil and dropper bottles for water to suspend specimens.
5. Lens paper for cleaning objectives, and Kim wipes for working with microscopic slides and specimens.
6: Bunsen burner, dissecting needle
Procedures:
1: Follow the instructions on the preparation of fungal spores for observation under the microscope.
A: Slide mounting
A slide was lie on bench and added a small drop of sterilized water. I opened the Bunsen burner and lighted it, the I sterilized the dissecting needle with burner to heat up. Petri dish with the culture was opened carefully right under the Bunsen burner, the sterilized dissecting needle was cool down by touching to the agar without fungus growth. The dissecting needle was gently touched the culture and transferred to slide with water. Covered the slide with cover slip, and gently knocked with the dissecting needle’s handle to squeeze out the air bubble.
B: Tape- mounts
An approximately length of Scotch tape was used to prepare the tape mounts. The stick side of tape was used to touch the fungus culture. All the procedures were performed under the Bunsen burner as described previously. Then the stick side of tape was stick to the slide, cover slip was not need in this preparation.
2: Setting the component microscope and the observation of specimens.
I was followed Dr. Shaw’s instruction to set up my microscope properly, and adjusted it before I placed my specimen.
Observations:
Alternaria brassicicola
Aspergillus niger
Cladosporium sp
Discussions:
Microscopy is an important skill for plant pathologists both for fundamental and applied studies of plant diseases. In this class, Dr. Shaw gave us a very detail description about the fundamentals of microscopes and the proper way to set up. We also gave the chance to learn the slide mounts and tape mounts. Under the microscopes we are able to clearly observe the structure of conidia or spore in different fungus as I listed before. I got a hard time to find the conidiophores, it may cause by the culture I picked is over-grown, and already formed the mature conidia or may be the techniques I used are hard to observe the intact conidiophores. I may think about to use other techniques to overcome this problem.
