Preparing a Wet Mount Microscopy Slide for Mushroom Spore, Mushroom Tissue Culture or Mushroom Mycelium Study
Using the microscope to observe and identify mushroom spores, tissue cultures or mycelium characteristics is an important step in the identification and taxonomy of mushroom species. The world of fungi is a fascinating one, full of unique and intriguing genera, each with its own structural characteristics and microscopic features.
Preparing a wet mount slide from pure, viable, high quality mushroom spores, mushroom tissue cultures, or mushroom mycelium is the first step in successful microscopy for mushroom identification and classification studies. Using professionally prepared spore syringes, tissue culture syringes or mushroom mycelium is an easy and convenient way to obtain pure microscopy samples for study. Sporepod.com carries a nice selection of spore microscopy kits for edible, medicinal and exotic mushroom species including: common button mushroom, portobello mushroom, oyster mushroom, shitake mushroom, reishi mushroom, psilocybe cubensis , psilocybe tampanensis, and psilocybe cyanescens. Once you obtain your spore, tissue culture or mycelium samples, you are ready to prepare your slides and begin your microscopy studies.
You will need to gather the following equipment:
- Microscope,
- Microscope slides,
- Microscope slide cover slips,
then follow these simple steps:
A. To prepare a wet mount slide for observation under a microscope:
- Place your microscope slide on a flat surface. Be sure that it is clean, dry and free from debris.
- Gently dispense 1 drop of your spore, tissue culture or mycelium sample from the syringe onto the slide.
- Grasping the coverslip gently by the edges with your forefinger and thumb. Place one edge of the coverslip near the edge of your sample on the slide. Gently lower the coverslip over the sample with the edges in line with the edges of the slide.
- If air bubbles form under the covers lip, gently press on the coverslip to remove them.
- An absorbent cloth or towel can be used to absorb excess water and flatten the wet mount slide by placing it at the edge of the coverslip where it can draw out some of the water.
B. Observe spores for identifying and unique characteristics: use 400x-1000x magnification:
- Determine whether spores are amyloid, dextrinoid, or inamlyoid by adding Melzer’s reagent to your slide sample. The iodine contained in Melzer’s Reagent will cause one of the following three color reactions which is visible if you hold the slide up to the light or under the microscope:
- bluish black = amyloid
- reddish brown = dextrinoid
- no noticeable change in color = inamyloid.
2. Observe for spore shape. Spore shapes generally fit into one of the following five categories:
Image: Kuo, M. (2006, February). Using a microscope: Viewing and measuring spores. Retrieved from the MushroomExpert.Com Web site: http://www.mushroomexpert.com/microscope_spores.html
- Round or nearly round (globose or subglobose)
- Tear shaped (lacrymoid)
- Elliptical or nearly elliptical (elliptical or ellipsoid)
- Football shaped (fusiform)
- Funky (diverse, asymmetrical, irregular shaped)
3. Measure spore size using the ruler in your microscope eyepiece. After taking your measurements, be sure to convert them to microns using the conversion multiplier established when calibrating your microscope. Generally, it is best to measure 10-12 spores to establish a measurement range in microns (ie.7-9.5u x 3-4.5u). It is also best to measure ornamented spores by measuring both the spore body and the ornamentation separately.
Image: Kuo, M. (2006, February). Using a microscope: Viewing and measuring spores. Retrieved from the MushroomExpert.Com Web site: http://www.mushroomexpert.com/microscope_spores.html
4. Observe spore surface characteristics. To view spore surface characteristics you will need to add Melzer’s reagent to your slide sample. Spore surface characteristics generally fall under one of the following eight categories:
Image: Kuo, M. (2006, February). Using a microscope: Viewing and measuring spores. Retrieved from the MushroomExpert.Com Web site: http://www.mushroomexpert.com/microscope_spores.html
- Pitted spores
- Smooth, dextrinoid spores
- Reticulate spores
- Smooth, inamyloid spores
- Warty spores
- Partially reticulate spores
- Amyloid, spiny spores
- Spiny spores (spinose, echinate, echinulate)
C. Observe mycelium for identifying and unique characteristics: use 400x-1000x magnification:
- Mycelium characteristics generally fall into one of the following categories for each form and color
FORM:
- Linear
- Rhizomorphic
- Cottony
- Zonate
- Matted or appressed
- Powdered
- Unique formations
COLOR:
- White
- Yellow/orange/pink
- Brown
- Blue
- Black
- Multicolored
The ‘Mushroom Cultivator” by Paul Stamets and J.S. Chilton contains detailed pictures and further descriptions of these unique mycelium characteristics. It is an excellent reference book and resource for microscopic mushroom studies.
D. Record your microscopy findings in your research journal and compare them to taxonomy charts for your specific strain or specimen.
With the right equipment and quality mushroom spores, tissue cultures or mycelium samples, you should have endless opportunity to study edible, medicinal and exotic mushrooms and successfully classify them according to genera, species and strain.
