How To Make Spore Syringes

Introduction

Spore syringes are the most common way spores are sold and used within the microscopy community. While their shelf life is shorter compared to those of spore swabs or prints, they’re extremely easy to use which also makes them great for beginners. In this blog post, we’ll be teaching you how you can make your own spore syringes at home!

A spore syringe is simply mushroom spores suspended in sterilized distilled water contained within a syringe. Each one can contain millions of spores! They differ from liquid cultures in a few ways. The first major difference is that spore syringes only contain mushroom spores while liquid cultures contain actual live mycelium. For this reason, liquid cultures are also known as “live cultures”. 

Second, spore syringes are filled with only sterilized distilled water while the liquid culture syringes contain some sort of nutrient source such as Light Malt Extract (LME), Karo, honey, etc. Because liquid cultures are considered to be “cultivating the mushroom”, vendors in the US are not able to offer active species in that medium. 

One benefit of spore syringes is that they’re better for storage than liquid cultures because they have a longer shelf life and can also be stored at more variable temperatures. Live cultures often need to be kept at specific temperatures to stay good. 

Most spore syringes come in a 10cc syringe, but if you make your own you can of course do more or less than that. When we make our syringes here at Sonoran Spores, we typically do them in batches of anywhere from 10-100 per batch depending on the popularity of the variety or species. With that being said, we use Pyrex Beakers to mix up our spore solution. These are great because they make it easy to measure how much spore solution you’re making, plus they’re easy to clean! But, if you’re making a small batch or even just 1 spore syringe, you can use a small mason jar or even a shot glass. 

Materials

• Spore print
• Distilled water
• Beaker, glass mason jar, or shot glass
  • If you’re making syringes of multiple species or varieties, then you’ll need one for each variety
• Pressure cooker that can reach 15 PSI
• 70-85% isopropyl alcohol
  • NOTE: DO NOT use anything higher than 90% isopropyl alcohol. It will evaporate too quickly to properly sterilize anything
• Scalpel handle and blade
• 2 mason jars
• Aluminum foil
• Magnetic stirrer and stirrer bar(s) (not needed if using a shot glass)
  • If you’re making syringes of multiple species or varieties, then you’ll need one magnetic stirrer bar for each variety
• Flame torch
• Paper towels
• Gloves
• Face mask
• Sterile and empty syringes
• Syringe caps
• Sterile environment

Tips Before Starting

• You should ONLY use distilled water to make spore syringes. DO NOT use tap or drinking water
• Having a sterile environment to actually make the syringes in is key if you want to have spore syringes that will provide you with clean, uncontaminated research. Working in front of a Laminar flow hood or HEPA filter is ideal, however you’ll still be able to make viable syringes using a still air box. Here at Sonoran Spores, we make all of our products in front of two flow hoods complete with HEPA filters to ensure the best and cleanest quality we can
• Something that’s important to note is that no matter how sterile the process is, it is always possible for spores to carry contaminants. The reason for this is because spores come directly from the mushroom which is in itself, not sterile. But, working in front of HEPA filters, or even in a still air box, as well as using properly sterilized and cleaned materials will greatly reduce the risk of contamination.

Prep & sterilization

1. Take your 2 mason jars, put the scalpel handle in one and syringe caps in the other and screw the lids on both jars. The amount of syringe caps needed is dependent on how many spore syringes you plan to make
   • TIP: Always add a little more caps than you intend to use just in case you spill or drop some while sucking up solution later
2. Decide on which kind of container you want to use to make the spore solution (beaker, mason jar, shot glass etc) and thoroughly wash it out using soap and water. Place them upside down on a clean cloth or paper towel to dry (upside down to keep any dust from getting inside)
3. Thoroughly wash the magnetic stirrer bars with soap and water. Place them on the clean cloth or paper towel to dry (if you’re using shot glasses, you can skip this step)
4. Put the magnetic stirrer bar in the beaker or jar and fill with the desired amount of distilled water. Cover with a layer of aluminum
   • If using a shot glass, pressure cook a mason jar of disstilled water (with foil on top). Youll sanitize the shot glass and pour the water in in your clean work area later. You will also not need to use a magnetic stirrer or stirrer bar
     • If you know your shot glass is made of thermal glass that can withstand the pressure cooker, then you can pressure cook it with the other materials. If it isn’t made of thermal glass or you’re unsure- DO NOT pressure cook it and sanitize as mentioned above
5. Use aluminum foil to completely cover the top of the beaker/mason jars of distilled water as well as the jars with the scalpel handle and the syringe caps
6. Put everything inside the pressure cooker and put the top on
7. Turn on the stove and bring the pressure cooker to 15 PSI for 45 minutes
   • NOTE: Do NOT start the 45 minute timer until the pressure cooker reaches 15 PSI. If it drops below 15 PSI, pause the timer and then resume it once the pressure returns to 15 PSI
8. Once the timer goes off, turn off the stove and allow everything to cool for 8-12 hours
   • NOTE: Once the stove is off, let the heat and pressure naturally release from the pressure cooker. Do NOT take off the pressure regulator or lid.

Making the Solution

NOTE: Anytime you’re working in your sterile environment and handling spores, you should be wearing gloves and a face mask. Be sure to use hand sanitizer after you touch anything non-sterile or right before you begin working with the spores

1. Gather all of your materials and the pressure cooker that has the scalpel handle, syringe caps, and distilled water
2. Start getting your sterile work environment prepared
   • If you’re using flow hoods, start running them 20-30 minutes before you start making the solution
   • If you’re using a still air box wipe the enire thing; box, lid, inside, and outside with isopropyl alcohol
3. Once you have your work area ready and your materials on hand, begin wiping down all surfaces and materials using isopropyl alcohol. Using the alcohol and a paper towel you will want to wipe down the following items:
   • Table/flat surface you’re working on
   • The bottle of isopropyl alcohol itself
   • Hand sanitizer bottle
   • Flame torch
   • Magnetic stirrer
   • if using a shot glass, sanitize it now
4. Remove everything from the pressure cooker and place it on the now sanitized table or in still air box
5. Remove the scalpel handle from the mason jar and attach a fresh blade. Flame sterilize the blade until it’s red hot and let cool for ~30 seconds
   • TIP: use the mason jar lid to rest your scalpel while it cools so that the blade does not touch the table
   • NOTE: You MUST make sure to let the blade cool because if it is too hot it will degrade or even kill the spores upon contact
6. Put the beaker of water on the magnetic stirrer and turn it on
   • TIP: You’ll see that when the magnetic stirrir is on, it will create a whirlpool of water in the beaker. You want the bottom point of the whirlpool to be close to the magnetic stirrer rod, but not touching it. If it’s too close or too far away, it will not properly mix the spores into the solution. To get closer to the stirrer rod, speed up the spinning. To get it farther, slow it down
   • If you are using a shot glass: Pour the distilled water from the mason jar. No stirring needed yet
7. Take your now cooled scalpel blade and scrape spores from your spore print
   • How much spores you need is dependent on many syringes you want to make. You can play around with the spore count to make it how you like. But, once you see spores spinning around throughout the solution then you have more than enough for microscopy research
   • If you are using a shot glass: Still scrape spores and dump them in
8. Once you have your spore solution mixed, pour out some syringe caps from the mason jar you pressure cooked them in. You only need to pour out enough for the amount of syringes you plan on making. It’s nice to pour a little extra though, in case you drop some along the way.
9. Remove the syringe from it’s packaging and use it to suck up your desired amount of  solution from the beaker
   • If you are using a shot glass: Suck up the solution and squirt it back into the shot glass. Do this 3-5 times or until the spores are adequately mixed throughout the solution. Once the solution is mixed, just suck up the solution again and move onto the next step
10. After the syringe is filled, close the syringe with a syringe cap
11. Continue until there is no more spore solution remaining in the beaker/shot glass
12. If you are doing more than 1 species/variety then sanitize your hands and workspace and repeat steps 5-10

Storage

• Spore syringes are best kept in the refrigerator and out of direct sunlight. If kept this way then they will remain viable for about 1 year. Although many people have reported getting great research results from spore syringes kept in the fridge that were multiple years old
• You can keep them at room temperature in a dry area, but it may shorten the shelf life
• The reason it’s best to keep them away from direct sunlight is because the UV rays can damage the spores if they are exposed for long periods of time
• DO NOT FREEZE