Printing ponderance

[Santiago]

I am always so nervous about making and using my own spore prints. I haven't done it yet, nor have I done agar work. The reason for this is because I don't have the space to really spread a project out, and everything has to be used quickly and then tucked away.

But I was thinking about this method for printing - let me know what y'all think.

Take a jar with 2 silicone ports and PC it empty. Then, in a glovebox, take a cap, open the jar, and put the cap in there. Wait some period of time (not sure on this - I just want to see if the procedure would work), then remove the cap. This should leave a spore print on the glass, right?

So then you should just be able to inject an airport, and then lots of sterile water, draw it up, and have a spore syringe.

Does this have a shot of working?

The biggest thing I could think of that would make it not work is that there would be no air flow in the jar while the cap is in there. Would a tyvek patch help this situation?

[retack]

i don't think airflow will be any concern here, only sterility.

also, i think it would be much easier to have the cap deposit spores onto foil that you could then fold/freeze/etc., instead of the glass itself which must be collected afterwards.

i'm nowhere near the stage of getting spore prints, but i'm intrigued at an idea i read about just injecting sterile water from a syringe into the gills of a cap, then sucking the water back in along with spores that it collects. well, i think that's how it went, it's been a while since i read it

[teesus]

youre idea is good, the only problem would be that the cap you put in the jar must be steril itself to not contam it. you can put the cap on a grill over the opend jar. the spores fall into the jar

[mestophilies]

teesus scored you right Santiago, get some small gage wire and make you a hanging basket from the top of the jar to hold your cap. It isn't really as scary taking prints as it seems to be. I feel you on the Agar work though, I really want to try it just a little too space confined.

[Santiago]

hey, that's a great idea!

ima try it.

[DjDizzy]

Originally Posted by Santiago

Take a jar with 2 silicone ports and PC it empty.

since it's empty - would putting the empty jar in the dishwasher be enough?
i know the advantage of a PC is the first word in the name, but does the pressure really matter in the sterilization process, wouldn't the water in a dishwasher be hot enough?

[crazy1]

you could also wipe the cap down with iso and make a wire hanger to suspend it in the jar. I"ve done this with success.

[Fresh Brewed]

This is sort of how my FOAF does prints. He uses silicon to make 3 supports around the inside of a 1/4 pint jar. He then makes a wire frame which fits into those silicon supports. Silicon ports in the lid and a few snips of stainless steel in the jar complete it. The printing jar then gets PCed. After collecting the print, the cap and wire frame get lifted out of the jar and the ported lid replaced. When it's time to use the print, a magnet is first used to manipulate the stainless steel from outside the jar to "scrape" loose the print. An easy way to go from print to syringe without a glove box.

[Looky]

The problem is going to be the spores will be stuck to the bottom of the glass and will not want to come off. This is the best way I have found to take prints. Cut the foil pieces and put them in a large tupperware. Then put in a little damprid (I put mine in one of those permanent coffee filters), then put the caps on the foil and close the lid. The damprid will help dry the caps so they don't deposit any moisture and it will also help force the caps to drop spores. Then after 12-24 hours take the caps out and reclose the lid. I like to let the foil sit in there with the damprid for another 12 -24 hours to make sure that there is no moisture and then put them into little baggies. They have perfect baggies for these at Michaels in the "making jewely" section.

[Santiago]

Lookyhere, I was planning on adding sterile water to the jar after printing, and then drawing up syringes. So I'm not too worried about getting the spores out of there.

DJDizzy - putting a jar in the dishwasher is much more of a hassle than just PCing it. Plus I can take my whole PC into my inoculation room and open it about 6 inches from the glovebox and put it in.
But I guess if you don't have a PC and wanted to try it, you could. I'd probably just steam a jar on the stove before I used the dishwasher though.

Crazy1 - 'iso the cap? Really? Wow, that sure would be awfully convenient, and rest my mind all the better. Would you do everything you could hit with the iso? That wouldn't result in some sort of freaky mutant spores, would it?

I'm going to engineer a little grate of sorts here pretty soon. I'll let y'all know what I come up with.

[Looky]

What I'm saying is, you are planning on getting the spores out of there by drawing them up in a syringe of spore solution right? I think that is was Hippie that said that the spores will stick to the glass and you won't be able to get most of them off of the glass. You will be drawing up mostly water without spores.

[Santiago]

I see. Yes, I suppose that makes sense.

But now I'm wondering if perhaps pp5 plastic might work better?? They'd probably stick to that too.

[eatyualive]

if your worried you can heat your oven to 350. put your foil cuts and glass ware that you place over the prints in the oven for 30 mins. take into clean area and allow to cool. then take prints when cool. make sure you have your sterile area setup prior to this.

if your having trouble building a glovebox or something for a clean room. a decently cheap thing to build is a shmuvbox.

[crazy1]

yeah they do stick to the glass..........trust me on that.
I really like to print on foil though
Seems to be the best way imho

[Looky]

In the discussion in a thread talking about this very thing, I saw mentioned, the possible use of a washer or nut/bolt added to the jar to be used to agitate and help break the spores from the glass. Foil it the way to go IMO.

[Fresh Brewed]

Originally Posted by Lookyhere

In the discussion in a thread talking about this very thing, I saw mentioned, the possible use of a washer or nut/bolt added to the jar to be used to agitate and help break the spores from the glass. Foil it the way to go IMO.

Stainless steel and a magnet and you are golden.

[sic]

Originally Posted by Santiago

Lookyhere, I was planning on adding sterile water to the jar after printing, and then drawing up syringes. So I'm not too worried about getting the spores out of there.

There's a video on youtube that is very helpful. SWIMS variation is using a half pint jar that's sterilized in a pc and then put inside of a glovebox. Prints are taken by using a sterile blade to cut and place 1 cap carefully into the bottom of the sterilized half pint jar. It's left to sporulate for 1-3 days with the half pint lid placed on or left slightly off inside the glovebox to allow for moisture to escape. After the print has been taken the lid can be put on the half pint to save the print.

When the print is ready to be used a sterilized exacto can carefully scrape the adhering spores from the bottom of the jar. Another half pint jar is filled 3/4 with distilled water, pc'd, cooled, and then added to draw spore solution. Often SWIM stores unused spore solution by placing the lid back on top of the half pint and placing the jar into the fridge.

SWIMS technique for reusing syringes: Unscrew the needle and placing all components in an ISO bath. Draws up 3-5 pulls of ISO with the needle off and then puts on the needle. Draws one pull with the needle on and flushes empty while spraying out the ISO into the needle cap. Put the needle cap on the syringe and wrap in foil. 1-3 wrapped syringes are put into an empty pint glass and pc'd on top shelf of PC.

[BuckarooBanzai]

Put a handful of marbles in the bottom of the jar, so they form a layer two or three marbles deep.

The spores will stick to the marbles, but you can shake the jar up (not violently - lots of marbles can break jars) and get them to mix.

Definitely PC the jar/marbles.

Definitely use a suspension source of some kind to keep the cap from touching the marbles.

Definitely do everything in the glove box.

When ready to use, just shoot 200-300 mL of LC into the jar, shake and incubate.

I did this at first but gave it up. Too much storage space hassle.

Printing on foil is really much easier than you would think.

[Santiago]

Just scared to try new things. If buying spores weren't such a hassle for me, I'd just do that again. I do have options though.

[BuckarooBanzai]

It is all easier than you think, man. Just read, read, read and move slow in the glovebox.

Be sure of what you are doing and why, you will be able to do it!

Remember, I had never even heard of cultivating mushrooms indoors 3 years ago. If I can figure this out, most anybody can - I ain't the sharpest spoon in the drawer!

[traskprater]

Well Santiago, I am glad you brought this up... because I have a question myself... Someone let me know if this step by step process would work:

1. sterilize a knife and cut the cap off
2. sterilize a jelly jar with rubbing alcohol (let dry)
3. use plain white typing paper to lay cap on
4. put the jelly jar upside so that it covers the cap and paper
5. wait 24 hours
6. remove jelly jar, remove cap
7. replace jelly jar over spore print
8. let dry for another 24 hours

[TastyBeverage]

Paper is generally not recommended for printing because spores can germinate on it and attempt to eat it as substrate. I see no issue with your process if you switch the paper out for a piece of foil though. Hmm, you may want to let the print dry for longer than 24 hours.

[Santiago]

Originally Posted by BuckarooBanzai

It is all easier than you think, man. Just read, read, read and move slow in the glovebox.

Be sure of what you are doing and why, you will be able to do it!

Remember, I had never even heard of cultivating mushrooms indoors 3 years ago. If I can figure this out, most anybody can - I ain't the sharpest spoon in the drawer!

I just don't see how a print could be sterile that comes off of a mushroom that's come off of a cake chillin' in an unsterile environment. How how how.

[BuckarooBanzai]

The foil is sterilized, by alchol or PC.

The spores are sterile when they fall, because of the way they grow. Before they fall from the shroom, they are in little sealed sacs, maturing.

As long as you work carefully, in a clean glovebox, very little other than spores will fall on the foil. The spores are sterile when they fall.

Once you have your technique down and have done it a few times, making clean prints is very easy.

[Santiago]

Okay BB, I trust you, but I'd like to just know WHY. Here's the thing - the mushroom is NOT sterile, right?? So how can the spores be sterile? It seems like some cross contaminant is bound to come from the mushroom.

I'm obviously overthinking. I always do this.

[BuckarooBanzai]

Mushroom spores grow inside microscopic sacs, almost like a tiny womb.

The sac is sealed until right before the spore drops, so no matter how dirty the area the mushroom grows, the spore is sterile right up until it drops.

[Dr_T]

You're overthinking, that's right. Try one or more methods, see what happens.

Don't know about anybody else, but I've had perfectly usable results just setting the cap on the foil. If you could suspend it, obviously that's cleaner.

I've wondered about suspending in a jar with water in the bottom- maybe they wouldn't stick to the glass that way. Maybe I'll try it some day.

[Santiago]

Haha. That WOULD be the next logical step. If they got on the sides, you'd probably still be screwed. But that's a good idea Dr T.

Ok ok. I'll be taking some prints soon.