Microscopy Guide

Studying mushroom spores under the microscope is a fascinating and rewarding hobby. You don’t need a full lab to get started, just the right kit and a clean setup.

We have put together simple step-by-step instructions to guide you in studying spores under a microscope. This is the process we use to validate the viability and cleanliness of our spore syringes before they are sold.

Essential Equipment

Sterility and Environment

• Still Air Box (SAB) or laminar flow hood
• Nitrile/latex gloves
• Surgical or FFP2 mask
• Paper towels
• Optional: Butane lighter or spirit lamp (for flaming syringe tip)

Preparing Your Workspace

Working under a laminar flow hood or in a clean room is the easiest way to keep all your procedures sterile, but we understand not many people have access to one of these options. The next best technique is to use a Still Air Box (SAB).

Always ensure you are wearing clean clothes and have clean hands. The cleaner everything is the better chance you will have for success.

Still Air Box Setup

It is essential to work in a SAB because dust and floating spores from the air can mess up your slide or give you false contamination results even when the spore solution is in fact clean.

• A Still Air Box (SAB) is a sealed, clear plastic container with two armholes. It has no airflow, which helps to prevent dust and spores from circulating.
• Wipe the inside of the SAB thoroughly with 70% alcohol before each use.
• Prepare your spore syringe by shaking it up to distribute the spores evenly. Any spores may get stuck in the needle if there are clumps present. A good way to distribute spores evenly is to place them on a vibrating surface (like your phone) for a few minutes.
• Give your materials a quick wipe with 70% alcohol and place them inside the SAB: spore syringe, slides, coverslips, and alcohol wipes.
• Wash your hands thoroughly and put on gloves.
• Spray/wipe your gloves with 70% alcohol before entering SAB.
• Work slowly inside the SAB to avoid air turbulence.

Slide Preparation

You’re only dealing with a very small drop of solution but this part matters. The goal is to get a clean, evenly spread sample under the coverslip, without air bubbles or flooding.

1. Shake the syringe to suspend spores evenly, they settle fast.
2. Remove the cap and attach a sterile needle.
3. Expel a tiny drop (about the size of a sesame seed) of spore solution directly onto the centre of a slide.
4. Replace the cap on the needle (don’t remove the needle). Next time you use the needle, ensure you flame sterilise it before use.
5. Hold the coverslip by the edge with sterile tweezers or gloved hands and lower it at a slight angle so it touches one edge of the droplet first, then gently let it fall flat.
6. Remove the prepared slide from the SAB carefully.
7. Label the slide to keep track of your work (species/date/source)

Microscope Setup and Viewing

This is where the magic happens, and where a lot of people get stuck. Here’s how to get a clear view of your spores without wasting time or wrecking your slide.

Step-by-Step Viewing Process

• Place the slide on the Microscope Stage with the coverslide facing up
• Start at 10X objective (low power). Turn on the light (not too bright), and use the course focus knob to bring the image into view. Move the slide left and right until you see the spore solution droplet. (You won’t see spores yet, just the droplet)
• Gently switch to 40X objective (medium power). Use the fine focus knob to sharpen the view. Spores should start to appear as scattered or overlapping, oval or round shapes.
• When you are confident, and you wish to see the finer details of the spores, you can switch to 100X objective with oil immersion. Place a single drop of immersion oil directly on the coverslip above the spore droplet, and rotate the oil lens into place, it must touch the oil. Focus very slowly with the fine knob only.

What should spores look like?

When you’re looking at spores under the microscope, you’re looking for at the shape, colour, texture, and condition. These details can help you confirm species, check quality, or spot contamination. Here’s what to expect (and what to question):

Psilocybe Species

• Shape: Typically elliptical (oval) to subrhomboid (rounded diamond shape), often slightly asymmetrical.
• Surface: Smooth and uniform, with no visible ornamentation.
• Colour: Medium to dark purplish-brown or brownish-grey, depending on lighting and maturity.
• Germ pore: Often visible as a slight flattening or light spot at one end of the spore. Only visible with 100X oil immersion.

Panaeolus Species

• Shape: More spherical or almond-shaped (sometimes slightly flattened at one end).
• Surface: Smooth, clean edges, and often slightly darker than Psilocybe.
• Colour: Jet black to blackish-grey , these spores absorb more light and appear heavier in the field of view.
• Clumping: Spores may cluster more tightly due to their darker pigmentation and smaller size.