|By Nan (Nanook) on Sunday, October 14, 2001 - 12:08 pm:|
SPORE PRINT TEK w/Providone-Iodine 10% Sanitizer
The ability to take a clean spore print assures the grower independance from outside suppliers. It is the first "Tek" a novice grower should attempt upon successful fruiting. A properly stored print has a documented shelf life of 5 years, and they are legal to sell and trade.
Let me tell you a story...
Fruiting on enriched cased straw a-la Nanook Straw Tek, A giant popped up to entertain the audience. It had a cap like nothing I had seen since I left Palenque years ago. It was a monster.
It was clustered in with five 20-30 gram shrooms. When the cap turned up and it began to drop spores, I realized my print teks were not what they needed to be. I had been printing in half pint jars with PF Tek but this cap dwarfed a 1/2 pint jar. I did what any respectable myco would do, ran to wally mart.
This Tek evolved while contemplating wally's wide selection of products targeted for todays mycophile. Wally did well...
I bought a large, clear glass, cheese cover (with plate) for about $8.00. I picked up a set of cylindrical plastic measuring cups for under a dollar. I am always about cheap (mushrooms should be free for everyone)... I got change from a $10.00 bill for a Sanitizable printing hood and mounts. Gotta love Wally
Warming plastic makes it easier to cut whether you are making cap mounts for printing, or cutting holes in storage containers to make a Glove Box. Warm the handles and bottoms of the plastic measuring cups with a blow drier, and remove them with a very sharp knife (I hate to say it but a box cutter is perfect). Out of the set of cups you want a nesting set of concentric cylinders... Rings that stack inside each other. These are Sanitizable mounts of varying sizes for printing caps. In this case the one cup measure was called for.
Preheat the oven to 300*F and turn it off. Sanitize the plastic ring and glass cheese set to "Glovebox Clean", using alcohol on a clean rag for the final wipe. Now alcohol and Lysol, most disinfectants in fact, are deadly, instantly lethal, to mycelium. Even the alcohol vapors will kill the mushroom gills and halt printing. I soaked some Heavy Duty Aluminum Foil in rubbing alcohol, lifted the cheese cover, set it inside, and placed the "print hood" in the oven to warm and dry off the disinfectant vapors. Caps need to print in clean, humid, air. Once the oven was cooled I was ready to print:
I picked the shroom with clean gloved hands, you can use plastic baggies right out of the box (they are sterile) for "pluckin gloves". I removed the stem from the cap with a pair of sanitized sizzors, cutting the stem off flush with the gills, and I centered the cap on the sanitized "one cup" mount. I worked on a fresh sheet of plastic wrap, so no contams contacted my plastic mount.
Now this is the controversial part. You can skip it:
I separated a two-ply white napkin, cut out a rough circle, and laid it over the cap. I mixed up a weak iodine sanitizing solution consisting of 10 drops of Betadine (generic equivalent: Providone-Iodine 10%) in 1/2 cup of Purified Water. I dropped enough solution on the napkin paper to wet it just to the point of dripping, then I trimmed the edges back flush with the cap using the re-sanitized sizzors. If you use rubbing alcohol to wipe your instruments, make sure they are flamed or completely dry before use.
I opened the oven, reached in and wafted off the cover of the cheese print hood. Wave it a bit under the cover of the oven to dispel all traces of lethal alcohol vapor. Your clean, cheesy print hood and Print Media are ready. Remember mycelia is also sensitive to Heat, make sure the oven and print hood are completely cooled before proceeding.
With clean gloved hands pick up the cap by the mount, center it on the printing media, and cover with the glass dome. Place it in an Incubator to print.
Replace the printing media every 12-24 hours. This also provides an air exchange to cap. If you tried the iodine sanitizer you may find the cap will print for 36 hours and give you three decent prints. This is what I found...
After 36 hours under glass my cap was starting to grow the dreaded White Fuzz!!! Don't Worry! Be Happy
Really. After 36 hours in a 1/2 pint jar at 87*F all of my other caps had stopped printing and had begun to curl and blacken at the edges. On the other hand this cap had white fuzz healing the severed stem, and rhizomes from the surface of the cap had permeated the napkin. The cap had transpired the water, indicated by condensation on the inside of the cheese cover. I kept it moist, wetting the cap, and exchanging air while swapping print media every 12 hours. It was still printing heavy, and to be honest, three large prints were plenty. But it was so outstanding... While the last print was dropping I made up and Sterilized a Dextrose Jar... Then I Cloned it too. Awesome shroom, that was months ago and it's still fresh daily ;)
Tips: Skip the napkin/Betadine if you are the nervous type or can't read directions. Full strength Betadine kills mycelia dead, if you don't measure and dilute I don't want to hear about your problems with this Tek. Betadine is NOT required to get a clean print. Your cap will be fine without Betadine until it curls, blackens, and dies. I think the Tek gives me an edge. You could also try just plain sterile water. Betadine has not been proven by a large number of people. My measurements could be wrong. You are on your own here, but be sure to report back ;)
P.S. Remember SANITIZERS Kill Spores!!!
I do not recommend straight tincture of Iodine on mycelia because it is just too strong. You can purchase Iodophor iodine sanitizer, it contains no alcohol and is perfect for this use. It is designed to be diluted. I use a few drops in 1/2 cup of water as a wash/dip solution prior to cloning and printing. Dip, set on a sterile plate for at least two minutes, you have dramatically reduced your surface contam count on your shroom tissue.
I have also used 10 drops of Betadine (generic equivalent: Providone-Iodine 10%) in 1/2 cup of purified (sterile is best) water as a wash.
Brewing & Food Industry Iodophor Sanitizer : Shroom Glossary
OK I was asked about this in chat... Sanitizing caps for printing. Here is what
I have worked out.
You want 25 ppm free iodine from either a dilute Betadine or generic Iodophor sanitizer.
25 ppm is approx 1 cc of iodophore in 500 cc water.
When sterilizing equipment for spore printing I place a glass with 250 cc of deionzed water and a cotton ball (cover with foil) in the pc. I also sterilize a foil wrapped pair of forceps.
Before I remove the mushroom cap for printing I uncap the glass with water and inject 1/2 cc of iodophore. I grab the wet cotton ball with the forceps, stir the sanitizer in, and use the cotton ball to wash the top of the cap after it is hooked and cut for printing.
Do not mess with the gills or turn the cap over for any reason. Wash only the top of the cap, gently, and allow excess sanitizer to drip off, then place in the mount for printing (some people place caps directly on the print media, I mount them in a wire basket or platform and suspend them just above the print media)
Printing caps need moisture to print well. One reason I like printing over water too is that the humidity stays high. You can also cover a printing cap with a sterile filter paper or thin cotton cloth (wrap materials in foil and sterilize in the pc). Use the cotton ball and forceps to moisten: stuff dries out fast (esp in a ventilated box). Apply sanitizer solution (be very careful not to drip), even directly to the top of the cap, in order to keep it clean and hydrated for extended printing.
A large healthy cap harvested at the beginning of the spore drop will print for days if it is properly set up and maintained and will produce several heavy prints