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YaMoonSun
The Double Standard


Registered: 10/23/14
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So I hear it's hard to print PE?
#22329641 - 10/03/15 06:16 PM (8 years, 3 months ago) |
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If I can't print it, I should still be able to empty a syringe of sterile water into a cap and quickly stir it up and pull it back into the syringe, no? Bad idea? Ever try it? Likely need to get a SAB for that procedure.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
#22329656 - 10/03/15 06:19 PM (8 years, 3 months ago) |
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Might work, why not sterilize some swabs and use those?
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YaMoonSun
The Double Standard


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Re: So I hear it's hard to print PE? [Re: Inocuole]
#22329658 - 10/03/15 06:20 PM (8 years, 3 months ago) |
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Have yet to find a store that carries agar, and telephone brand agar went up in price on amazon, and I'm not paying $6 shipping.
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mushpunx
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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
#22329662 - 10/03/15 06:21 PM (8 years, 3 months ago) |
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You'd be better off using a sterile swab under the gills if you want spores, or take a tissue biopsy and clone it. You would need agar for both, either to swipe the spores onto or for the clone.
I doubt the procedure you are describing would be clean enough for a clean spore syringe. Im not sure how most spore vendors get thier PE syringes, maybe they have a strain that produces enough spores to print?
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 Amateur Mycologists United AMU Q&A
Edited by mushpunx (10/03/15 06:24 PM)
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YaMoonSun
The Double Standard


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Re: So I hear it's hard to print PE? [Re: mushpunx]
#22329705 - 10/03/15 06:36 PM (8 years, 3 months ago) |
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Isn't thee idea to get the syringe as dark as possible (at least in this procedure) using clean/sterile water? Fungi only survive in nature because their vast amount of spores out-numbers their competition, right? I could maybe even avoid the SAB if I inject the sterile water through the unbroken veil and just give it a little shake, and then angle the mushroom to pull back all the spore solution back into the syringe. Presuming that the mushroom is mature enough to have even produced said spores.
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Grey
⇜ ✯ ⇝



Registered: 11/06/14
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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
#22329737 - 10/03/15 06:44 PM (8 years, 3 months ago) |
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Do you have a PC man? If you have one and want to take the dive into agar, I'll send you some. You did right by that print I sent you, I wouldn't hesitate.
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Fantastic Mr. Fox
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Re: So I hear it's hard to print PE? [Re: mushpunx]
#22329738 - 10/03/15 06:44 PM (8 years, 3 months ago) |
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Quote:
mushpunx said: Im not sure how most spore vendors get thier PE syringes, maybe they have a strain that produces enough spores to print?
I've been wondering this myself..
-------------------- Giving is all we have, for we're just grateful to be alive

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Inocuole
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Registered: 11/21/11
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Workman uses a centrifuge to separate the spores from cap material and contams.
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Grey
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Here you go guys. Credit to Blackout for the quotes.
Quote:
blackout said:
Quote:
Workman said: It isn't really a secret but the basic method is to take very mature caps that are visibly producing spores. Even these rarely drop decent prints but the spores are there. These are gently dried, which tends to cause even more spores to mature before the cap completely dries out. The caps are then crushed, rehydrated in sterile water with a wetting agent and then centrifuged. Spores are very dense compared to the mushroom tissue, bacteria and other impurities, so they separate nicely into a pellet at the bottom of the vial. Remove the top layer and repeat with a few more washings and centrifuging and you have nearly pure spores when you are done. This is dried and used as needed.
It is somewhat complicated and not practical for the home experimenter but it does scale up well for the production of commercial quantities of PE spores. This method can also be used with regular cubensis caps after normal printing. There are a surprisingly large amount of spores clinging to gills. Swabbing is easier and works just as well for archiving spore samples.
Quote:
Workman said: Its an adaptation of an existing method. I forget where I first read about it but I want to say Mycelium Running by Stamets. I'd have to confirm that. The method is self cleaning since the contaminates are lighter than the spores and deposit on the top of the pellet.
Quote:
Workman said: I left out a step. You should coarsely filter the mushroom material before centrifuging. Its probably obvious that you don't need large chunks in the centrifuge.
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AMU Q&A If you don't have a plan of your own, you'll become a part of somebody else's.
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YaMoonSun
The Double Standard


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Re: So I hear it's hard to print PE? [Re: Grey]
#22329762 - 10/03/15 06:50 PM (8 years, 3 months ago) |
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I still have most of it, thanks again  No pressure cooker yet, no. I just extend the steam times of everything I do based on size and educated guesses. Not in any rush when it comes to sterilizing. Just need to done  If you wanna send out some agar I'd appreciate it, but I can't make use of it until I get some micropore tape and maybe some little containers. How long is the self life?
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Inocuole
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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
#22329770 - 10/03/15 06:53 PM (8 years, 3 months ago) |
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Agar powder doesn't seem to have an actual... shelf life that I can tell. I don't know what would happen to it if it goes bad, I suppose it would have to stop being agar to go bad. Not sure if that's what he's offering though.
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YaMoonSun
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Re: So I hear it's hard to print PE? [Re: Inocuole]
#22329778 - 10/03/15 06:54 PM (8 years, 3 months ago) |
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That's good to know, cheers
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Fantastic Mr. Fox
Grass Seed Connoisseur


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Re: So I hear it's hard to print PE? [Re: Grey]
#22329792 - 10/03/15 06:59 PM (8 years, 3 months ago) |
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Yas! I know what Im adding to the Christmas list this yr. heh,
Thanks for citing, Grey!
-------------------- Giving is all we have, for we're just grateful to be alive

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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
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Quote:
Fantastic Mr. Fox said:
Quote:
mushpunx said: Im not sure how most spore vendors get thier PE syringes, maybe they have a strain that produces enough spores to print?
I've been wondering this myself..
I can tell you how I do it I find turning a swab from a user into multiple syringes is the best way to drag it out, and make it last.
Really it's so simple. Take sterilized water and pour into a baggie. Take swab and bring it into the baggie. With the swab in the water, massage the swab with your fingers from outside the bag. Draw the syringes. Horray!
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YaMoonSun
The Double Standard


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Re: So I hear it's hard to print PE? [Re: Grey]
#22329840 - 10/03/15 07:11 PM (8 years, 3 months ago) |
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mushpunx
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Re: So I hear it's hard to print PE? [Re: Mad Season]
#22329860 - 10/03/15 07:19 PM (8 years, 3 months ago) |
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That is so cool thanks for that!
I don't think its really worth it to make spore syringes with PE anyways past a vendor capacity.
If you are determined to do cakes you can always go agar to LI or LC.
You can slant a culture and be good on PE for life
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 Amateur Mycologists United AMU Q&A
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YaMoonSun
The Double Standard


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Re: So I hear it's hard to print PE? [Re: mushpunx]
#22329877 - 10/03/15 07:23 PM (8 years, 3 months ago) |
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Would likely save me some time. I have two LC maturing at the moment. What's the best way to get all the myc into the syringe? I some times mix the culture before I pull it into the syringe, and I haven't had any luck that way as of yet. Should I submerge the syringe in the culture to get the mycelium from the bottom of the jars?
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mushpunx
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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
#22329889 - 10/03/15 07:26 PM (8 years, 3 months ago) |
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Really man you shouldn't be be doing LC. If you don't have agar
Very risky!
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Mad Season
hookers and blackjack



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Re: So I hear it's hard to print PE? [Re: mushpunx]
#22329896 - 10/03/15 07:27 PM (8 years, 3 months ago) |
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Yeah definitely. But I did it because there was this 1 swab i got from someone I had a bitch of a time getting things to germinate. It would contaminate a lot too. I ended up hydrating it in 3 syringes and diluting it a shit ton. It worked much better. Much less contams, and the spores were able to germinate.
I've just weighed out the pros and cons, and if it's a swab i want to get some clean growth out of, no matter what, I don't like risking it on the few plates I'd swipe. Especially if they all turn bacterial.
If it's your own fruits hell yeah no need for any syringes. I just like the security of having more to fall back on in case it all goes to shit. Na'mean?
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YaMoonSun
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Re: So I hear it's hard to print PE? [Re: mushpunx]
#22329920 - 10/03/15 07:32 PM (8 years, 3 months ago) |
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How so? I have growth in both of them. Aside from bacteria
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micro
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Re: So I hear it's hard to print PE? [Re: mushpunx]
#22329930 - 10/03/15 07:35 PM (8 years, 3 months ago) |
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I would say you shouldn't do LC without something to sterilize it.
If you get septum-top bottles though, it's pretty easy to be sterile once you can PC the media.
If you need sterile spores you could always try fruiting it in a bag...
Does PE not produce them at all, or just very few?
Very few is okay but then you would certainly want agar since inoculating substrate with that would be painfully slow.
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Fantastic Mr. Fox
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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
#22329957 - 10/03/15 07:42 PM (8 years, 3 months ago) |
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Just because you see growth doesn't mean it's clean growth.
-------------------- Giving is all we have, for we're just grateful to be alive

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Grey
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Re: So I hear it's hard to print PE? [Re: micro]
#22329967 - 10/03/15 07:45 PM (8 years, 3 months ago) |
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They produce them, they just don't drop them.
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YaMoonSun
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Amen, but I'd like to take my chances. Worst possible outcome?
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cronicr



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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
#22330417 - 10/03/15 09:38 PM (8 years, 3 months ago) |
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Quote:
YaMoonSun said: If I can't print it, I should still be able to empty a syringe of sterile water into a cap and quickly stir it up and pull it back into the syringe, no? Bad idea? Ever try it? Likely need to get a SAB for that procedure. 
it has worked out before, i'd expect a little failure along the way though(should have a sab for everything)
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invitro

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Re: So I hear it's hard to print PE? [Re: YaMoonSun]
#22330675 - 10/03/15 10:40 PM (8 years, 3 months ago) |
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good info Grey, thx
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Violet



Registered: 12/06/11
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Re: So I hear it's hard to print PE? [Re: invitro]
#22331287 - 10/04/15 03:25 AM (8 years, 3 months ago) |
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That's cool, about the centrifuge.
I have used the syringe drops method to harvest PE spores before into a syringe. I found it more tedious than using a swab but equally, possibly more, rewarding. Thing is the droplet's surface tension stops it from easily getting down into the gills. And when it does, it's not coming back out. So what I did was: 1. Sterilized a syringe with 5-8 mL of water in it, with the needle attached, all wrapped in foil. 2. Used my extra mature and wrinkly PE fruit that had grown invitro - picked in my sterile airflow work space, and readied the syringe in my other hand. 3. Placed a few droplets of water on the gills while holding the fruit upside down. 4. Worked the water into the gills with the tip of the needle. Repeated 3 and 4 until excess water that could easily be sucked up was building up in the cap. 5. Without absolutely shredding the gills, ran the needle tip along the gills, with the goal to rub off spores into the water. 6. Sucked as much of the water up as I could.
If you feel good about having this all kept clean during the process, which I did in my sterile airflow, you can shake the syringe and put water back in to stir it with the water that remained from the first time and try to get more of the spores that you freed but were left behind.
To me, this was worth it. Because with 1mL of spore solution I can start like 10 of my grain petris for my culture tech.
-------------------- Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it! PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers The simplest, quickest, safest tek! For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers! • Violet's Teks and Posts •
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YaMoonSun
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Re: So I hear it's hard to print PE? [Re: Violet]
#22331992 - 10/04/15 10:05 AM (8 years, 3 months ago) |
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