By Hippie3 (Hippie3) on Saturday, May 26, 2001 - 08:45 pm: Edit

I use a simple method that's worked well for me. I use aluminum foil. I cut a 6" wide strip off the end of a roll, then fold it over in half. That give you about 3" by 12". I then put a cap every 3-4 inches, depending on the size. Then, the top lays over the caps, and the whole thing is stored in a small airtight box for 24 hours. Remove the caps, cut the foil into individual prints, then fold the edge of the foil over making a seal all around the print. Use a "sharpie" to label the print. Minimal work, and if the caps are from cultivated fruits and clean the prints come out clean. Foil is nice bacause you can scrape all the spores off it, I prefer it to paper, although wax paper is ok, but you have to tape the edges, whereas foil can be "self-sealing".

By Hippie3 (Hippie3) on Saturday, May 26, 2001 - 08:47 pm: Edit

Materials:Do I use typing paper, wax papar, aluminum foil, 3x5 card, microscope slide, cow tongue?

wax paper and aluminium foil works good, typing paper works also but a lot of spores are absorbed by the paper

Should I use tape, or shoudn't I?

use tape

Storage:Cool and dry, cool and damp, hot and dry, hot and damp? Stack verticaly, or file horizontaly? Plastic bags, or no?

Cool and dry, plastic bags..

Time:Leave the cap for one day, or one week?
one day

Cataloging:Just how many strains ARE there realy?
a lot of strains...comercially available something like 20-25, in the wild probaly thousands...

Maintinance:Re-print each strain every 1 year, 5 years, 10 years?

every 5 years if well stored

By Hippie3 (Hippie3) on Saturday, May 26, 2001 - 08:47 pm: Edit

I prefer to print directly into a sterile .5 pint jar,then you just gotta add sterile h20, stir,draw,and innoculate. use a sterile probe to place the cap in the jar,put the lid on ,wait till the spores pop out,then use the sterile probe to remove the cap,then reseal the jar.I do it all in the glovebox,and dont remove the jar till the process is complete. a wire stand (sterile)can be placed in the jar to keep the cap off the bottom,if desired

By Admin (Admin) on Saturday, May 26, 2001 - 10:57 pm: Edit

SPORE PRINTING AND SPORE SYRINGE PREPARATION

The mature specimens are good for spore production, but are not as good for consumption (weaker potency). They are characterized as becoming darker, with dark bluish colors appearing on the caps and stems. The cap upturns and reveals gills darkening a deep brown color. The mushroom will look like an umbrella that has turned up edges. On the stem can be seen the purple deposits of the dropping spores. Mature adult mushrooms release spores by the millions. In the area around the mushrooms can be seen a deepening color of purple. As the spores fall and collect they will color deep purple. This is the signal that the mushroom has matured and is now in its sporulation cycle. This is the time to take their spores.


SPORE PRINTING EQUIPMENT
KERR 1/2 PINT WIDE MOUTH (LOW FORM) CANNING JAR. (ANY SUITABLE JAR IS OK)
FINGER NAIL CUTICLE SCISSORS - (cosmetics - drug stores)
ALCOHOL, TEQUILA SHOT GLASS AND EYE DROPPER.
1. Presterilize the jar and regular metal lid (rubber edge up) in a small toaster oven at around 300 degrees Fahrenheit for around a half hour. Keep the lid loose during the sterilization cycle. When the jar has cooled down, tighten the lid until it is time to use the jar for a spore print. The rubberized edge will be a bit melted, but that won't be any problem in this technique.

Note: What follows is a sterile technique. The first rule that must be always followed is to wash hands prior to sterile work. Hands are a prime source for bacteria and microspore contaminants. Sterilize all the work surfaces with rubbing alcohol. Minimize drafts. Try for a still air environment. Don't breathe on the work. Run a small home appliance style HEPA air cleaner (99.97% rated efficiency - available at drug and department stores) for a few hours in a closed room to clean the air before doing sterile work.

2. Flame sterilize the scissors with an alcohol flame and snip off the mushroom cap. Cut the top of the stem as far up into the cap as possible so that the gills of the mushroom will sit flat on the surface of the jar bottom. With quick and sure movements, place the cap into the jar and place the lid on loosely. Pierce the top of the cap with a straight pin to pick it up and handle it.

3. Leave the jar with a loose cap for a couple of days in a draft free area away from direct sunlight. After the print is taken, quickly and with as little air disturbance as possible, remove the jar cap and extract the mushroom cap from the jar. With a loose jar cap, let the jar sit in a draft free place to dehumidify for a few days before sealing it up (with tape) because there will be some residual moisture left behind on the spores and glass. Store the spore print jar at room temperatures in a dark place away from sunlight. Don't store it in a refrigerator.

Psilocybe Cubensis spores begin to degrade a few months after they are taken. After approximately 1 1/2 years, spore germination will be greatly reduced or won't occur at all. Germination is massive and quick when the spores are fresh.


MAKING A SPORE SYRINGE
Materials list:

1. Spore print in jar.
2. Sterile syringe with water for injecting water into the spore print jar.
3. Sterile syringe for loading spore solution out of the jar.
4. A small Pyrex glass stirring rod (science - lab supply).
5. Alcohol, tequila shot glass and eye dropper.
6. Lid with two holes. Prepare this lid by drilling a hole in the center of the lid to fit the Pyrex glass stirring rod. Punch the second hole near the edge of the lid (rubberized edge up) to fit a syringe needle.


Syringe preparation
Boil a pot of water. Draw boiling water into a syringe and squirt it out several times. Refill the syringe with boiling water, replace the needle quard and wrap the syringe in tin foil. Prepare several syringes like this. Drop the syringes into the boiling water and boil them for one hour. Let them cool before using.
The main point of this technique would be to expose the interior of the jar to as little room air as possible. Always protect the holes in the lid by placing tin foil or sterile surgical tape over the holes before and after this procedure.



1. Inject sterile water into the spore print jar through the needle hole.
2. Flame sterilize the glass stirring rod and let it cool a minute. Insert it through the center lid hole and with the rod end, scrape spores into the water.
3. Insert the sterile syringe needle through the small hole at the edge of the lid. Tilt the jar until the water comes up to the needle tip and draw the spore water into the sterile syringe.

Store the syringe at cool temperatures in the dark. A properly prepared spore syringe will be good for several months and even up to a year or more.


PF homepage - www.fanaticus.com

By Admin (Admin) on Saturday, May 26, 2001 - 10:59 pm: Edit

How to Make a Spore Syringe

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Sterility is key. If your spore print is contaminated or you introduce contamination into the spore syringe, you will have difficulty later in the process. Ideally, there should be no fans blowing or drafts of air. You should clean the area where you will be working carefully and make sure that everything is tidy.
Materials needed:

spore print
10 cc. syringe with long needle Where to find
shot glass
regular glass
coffee mug
cigarette lighter
X-Acto knife or sharp steak knife
microwave oven
Several things need to be accomplished. First, we need to sterilize a shot glass to easily mix the spore solution and we need to sterilize a syringe to hold the solution. We also need some sterile water in which to suspend the spores. The following procedure will accomplish all of this.

Fill a coffee mug with water and place a shot glass inside the coffee mug. Make sure the shot glass is completely submerged. Place the coffee mug in the microwave oven and get the water to a full boil for 10 minutes. It does not need to be a violent boil. Adjust the heat level of the microwave oven to keep too much water from being lost if necessary.

Remove the shot glass and empty the excess water out of the shot glass. Place another glass over the shot glass. This will keep air born contaminates from settling in the shot glass while you wait for the shot glass and water in the spore syringe to cool.

Fill the syringe with hot water from the mug. Eject the hot water and repeat several times. This will insure the inside of the syringe and the needle are clean and sterile. This is especially important if you are using a syringe from a previous crop. When the needle is inserted into the substrate, it is possible to get nutrients up inside the needle and for contamination to grow. The last time you fill the syringe with hot water, do not purge it. Let it sit in the syringe until it is cool. This is useful for two reasons. First, the continued heat from the water can still work to eliminate any remaining contaminates. Secondly, once the water is cool it can be used as the sterile water needed to fill the syringe. Make certain that nothing touches the needle of the syringe.

The Psilocybe mushroom spores will be killed if they come in contact with anything too hot. You need to wait until the shot glass and spore syringe are at room temperature. When it is safe to proceed, use the cigarette lighter to flame sterilize the X-Acto knife and the needle of the syringe. Let the blade of the knife cool, but make sure it does not touch anything. When it is cool, carefully open the spore print and scrape a fleck of spores into the shot glass. A fleck 1/4 inch by 1/4 inch is more than sufficient for a 10 cc. spore syringe. Use the needle of the syringe to stir the spores into a few drops of water. Usually, there will be a few drops left over in the shot glass from when you emptied it. Otherwise, you can get the drops from the syringe. Stir the fleck of spores until they are well broken up and 'dissolved' into the water. Purge the water out of the syringe into the shot glass. Pull the water back into the syringe, being sure to suck the spores in also. Do this once or twice more to make sure the spores are well mixed in the spore syringe. Often, it takes several tries to get the spores fully broken apart and well mixed.

If the spores in the print have been dried and are not fresh, it is best to wait six hours to use the spore syringe. The spores need to rehydrate. If your in a hurry, the spores can still rehydrate in the culture jars.


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How to Make a Spore Print

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Once you have a mature mushroom, you are in a position to make a spore print and use it to continue cultivation of mushrooms. The cap should be harvested when the mushroom cap has become flat or is starting to invert.
Sterility is key. Adaptation-25 Be careful not to do anything that will compromise the sterility of your spore print. The typical procedure is to cut the stalk off of a mature mushroom very close to the cap. A sterilized knife or razor blade is used to do this. The cap is then laid on a sterile piece of tissue paper or card stock and a small glass set over it. The glass is needed for two reasons. First, it keeps the spore print insulated from airborne contaminates. Secondly, it helps keep the humidity high so the mushroom cap can continue to live and drop its spores. One note of caution. Some humidity usually needs to be allowed to escape. You want the environment inside the glass to be slightly less humid than the environment in which the mushroom was grown. If you have problems getting a cap to drop its spores, try using a piece of paper for the print that fits entirely inside the glass and spreading out a wash cloth flat on the table. Let the edge of the glass seal to the wash cloth instead of the paper. This will usually allow enough humidity to escape to cause the cap to drop spores.

If everything goes well, after a day or two the cap will drop its spores. Adaptation-26 There will be a purple-brown dust underneath the cap. These are the spores.




Eye glass lens paper is good source of sterile tissue paper. A box of waxy tissue paper that deli's use to pick up donuts and rolls is another excellent source of sterile tissue paper. Card stock (such as a recipe card) is a bit easier to use later when you want to prepare a spore syringe, but you have to expend the extra effort up front to sterilize it. To use card stock, place in a 425 degree F. oven for 15 minutes. Let it cool with a glass over it before you place the cap on it.
Once you have a spore print on the paper, remove the glass and cap. Fold the spore print in half and seal the edges so air can not get in. A piece of scotch tape on each side will do nicely. The spores will stay viable for 18 months if they are kept in a cool, dry and dark spot. If you place a small amount of desiccant in the bottom of a film container and place a cotton ball on top of the desiccant, you have an ideal container to keep the spore print. The cotton ball will keep the desiccant from touching the spore print. Seal the spore print in the canister and place the canister in your refrigerator until you are ready to use it.

Note that if you want to be self sufficient, it is a good idea to have multiple spore prints and store them separately. You just never know when you are going to be surprised with a massive contamination problem or thermal death. The safest thing to do is have a couple viable spore prints so it is easier to recover from disaster. A spore print is typically viable for about a year if it is stored in a cool, dark location. As a print ages, germination gets slower and this process becomes more prone to contamination.


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Combination of Spore Print and Preparation of Inoculant

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This excellent idea is from: [email protected]
If it is your intent to use a mushroom to immediately generate inoculant for a successive crop, you can eliminate some of the above steps and reduce the risk of contamination.

Poke a small nail hole in the cap of a wide mouth jar. Cover the nail hole with a piece of electrical tape. A 1/2 pint canning jar similar to that used for the cultures is fine. Sterilize both the jar and the lid.

Place the harvested cap in the sterilized, wide mouth jar. Seal the top with the lid and wait until a spore print has been generated on the bottom of the glass. Open the jar and remove the mushroom cap. Add 3/4 cup of sterilized water. Seal the top of the jar with the lid and shake the jar. The spores need to be mixed well into the water. This procedure elements the need to transfer spores from a print to some container to make the solution. The spore print is generated inside the container and the only extra step is adding water. It also has the benefit of making a large amount of solution that is easy to use.

Now, any time you need inoculant, you can shake the jar and peel the tape back to expose the nail hole in the lid. Simply insert the needle of the syringe into the inoculant and pull some into the syringe. The syringe needs to be sterile or you risk contaminating the entire jar of solution. The solution will keep longer if you refrigerate it. You still should keep some spore prints on paper because it is possible to contaminate the entire jar of inoculant if you make a mistake.