There are many, many, ways to print, some more, some less sterile. These are a few ways. Not the best or only ways, just ones Ive used successfully multiple times.
Printing tek(s). With Alternatives marked *A
Three methods listed will be, an "Oven method", a "Print Box" method, and "Simple method". All will use open face prints in mini zip bags. None use a glove box or hepa/laminar flow.
Some steps are universal. marked with *U ( info )
The biggest components of all. :
Be quick, prepared, move smoothly and precisely and most importantly, make sure every item you are using, and yourself (hands,gloves, clothes lighter, xacto,etc) are clean and sanitized! This is a constant.
Unless otherwise stated, assume this is law throughout every step , every method!
Items needed: all included but not needed for each method ( you can figure it out)
Aluminum foil
Iso alcohol 90% if possible, 70% will work
Exacto knife
Print box- a flat bottomed plastic food grade storage container with lid (gladware , tupperware, Sterilte etc)
A baking dish (size is dependent on how many prints you are making for both dish or box methods)
*Mini zips- 2x3" or similar. big enough to easily accept the foil squares. wlmrt craft aisle 50-100 for $1
*A Snack size zip bags
Jar, glass, plate, or just a clean flat disinfected surface (10% bleach/water solution)
Nitrile gloves (latex or vinyl work too)
Tweezers
The Simple Method-- *U1 -( Choose an open and flattening cap from a standing specimen. Wipe the top of cap with a folded paper towel moistened with Iso. (moistened, not dripping) Gently grasping the cap with one hand(see *A2), and using flame sterilized and iso wiped Exacto or razor in the other, cut the cap from a standing mushroom just below, but not touching the gills.
A tiny bit of stem stub is ok, but you want to cut as close to the gills as you can get without touching.
Place on a square of shiny side up, iso wiped, aluminum foil, slightly larger than the cap. This should be already prepared and waiting )--- on a clean plate or flat surface covered by small jar or glass. Immediately cover with the glass, or jar. Leave for 12-24 hrs. Check for spore deposit by slightly tilting the glass/jar and gently lifting the cap edge using the exacto. If not dark enough leave another 8-12 hrs and then it should be ready.
( Prepare your mini zip with an Iso wipe on outside only. Using one had crack the seal of the zip with a sliding motion.Lay the zip in a position close to the print jar. Quickly but with as little air disturbance as possible, remove the cap from the foil and set aside for drying. Remove the print using sanitized/ iso dipped or wiped tweezers, grasping only the edge of foil. With one hand and using the sliding motion, further open the zip ,while quickly inserting the foil/print with the tweezers . Zip it closed.)
That's it. Label the zip with variety/date etc. Its ready for making syringes or to place in an envelope, and put in a dark location for later use.
*A2- use a small sharp object to pierce the top of the cap just deep enough to hold it, instead of grasping with your gloves. This is for removal and transfer to and from the foil. A small flame sterilzed tack, pin, needle,fish hook or piece of wire etc.. It can be left in while printing( as long as it is sanitized/sterile and fits in the covering jar/glass) Makes it easy if you have one for for each print. A little bend or hook shape on the wire can help grasp the cap. The little "handle" is really handy.
*U2 ( All of the steps you should be using the gloves, sanitizing as needed with iso. Have your items ready, dont reach into your pocket in the middle to get your lighter for example. Have it prepped, iso wiped and within reach to flame sterilize the Exacto blade.
All of steps in every process are done in a manner to limit exposure to open air.)
I suggests practicing the method of opening the zips with one hand and inserting a blank foil square using the tweezers with the other. Also the "handle", practice on a cap you are not going to print, make sure it's secures the cap and doesnt fall off during transfers. In fact a dry run for all steps helps get the movement, process and helps lessen exposure time.
I will add the next method later for multiple caps in one container "Print Box" style, My preferred method, I will post a pick of what I print in when doing a large number. The size of the container is important in that the caps require a certain amount of humidity maintained to release the spore. A container with a totally air tight sealed lid can invite mold on the cap if left too long.
peace, jpc
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