Paradox
©
Fisana

Jump to content


Photo
- - - - -

How do you get a perfectly clean spore print.


  • Please log in to reply
45 replies to this topic

#1 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 15 February 2017 - 05:07 PM

My house is pretty much a dirty disgusting mold iinfected tear down.  With my finances I'm probably not moving right away.

 

I really don't wanna half to keep buying syringes because I can't complete the process of getting a clean print and putting it into production.  I've got a glove box which works ok for doing petri plates but it seems like an awful slow process.  I'd like to start doing straight spore to LC.

 

How do I get clean prints in this dirty environment?

 

Thanks.


  • fuzzyfunguy and tailsmcsnails like this

#2 CatsAndBats

CatsAndBats

    [^._.^]ノ彡 & /|\( ;,;)/|\

  • OG VIP
  • 9,227 posts

Awards Bar:

Posted 16 February 2017 - 09:34 AM

There are mountains of information here at mycotopia already, I'd start here:

 

http://archives.myco...html?1061991123

 

Are we talking cubensis?

Are you trying to create a spore library or are you just trying not to spend more money? If you just want more inoculant purrhaps just 'milk' already colonized jars:

https://mycotopia.ne...ing-super-inoc/

 

Do you have an SAB/GB/flow hood in your employ? That will solve almost all of your dirty house problems, that and agar is your friend.


  • MLBjammer, OysterFarmer and Heirloom like this

#3 dead_diver

dead_diver

    Stoned Since '76

  • OG VIP
  • 3,504 posts

Awards Bar:

Posted 16 February 2017 - 10:27 AM

Perfectly clean as in sterile? Very difficult to do but not impossible, just impractical.
Any small space like a closet that you can Lysol and bleach bomb should be reasonably clean. Make sure you are clean too.
  • OysterFarmer, Heirloom and CatsAndBats like this

#4 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 17 February 2017 - 06:22 PM

There are mountains of information here at mycotopia already, I'd start here:

 

http://archives.myco...html?1061991123

 

Are we talking cubensis?

Are you trying to create a spore library or are you just trying not to spend more money? If you just want more inoculant purrhaps just 'milk' already colonized jars:

https://mycotopia.ne...ing-super-inoc/

 

Do you have an SAB/GB/flow hood in your employ? That will solve almost all of your dirty house problems, that and agar is your friend.

Trying not to keep spending money.  Plus I'm sure they have some sort of record somewhere of all the times you order this shit.  Once or twice they figure you are just some curious hippy.  Do it again and again they probably look closer.

 

I just had a really good method.  Order syringe.  Put in a lc as soon as I open the box.  Use that to innoculate grain.  Went on like gangbusters but now trying to get the clean spores to work with just isn't going nearly as fast.  Its like ten percent as fast as it was.

 

I've considered maybe injection some liquid into a petri plate as I use small mason jars with holes in the lids but my liquids haven't been doing as well.

 

Long term though I do want to not only have libraries but be able to walk in the woods and grab a mushroom then put it into culture.  It would be impossible to farm the local mushrooms here by going through any distributor. 



#5 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 17 February 2017 - 06:30 PM

Perfectly clean as in sterile? Very difficult to do but not impossible, just impractical.
Any small space like a closet that you can Lysol and bleach bomb should be reasonably clean. Make sure you are clean too.

 

Well at least as sterile as the shit I've been ordering.  Haven't had any issues with my supplier in the years I've been using them.  But it just seems like a hole that should be closed needing to keep ordering.

 

I'm using a glove box at the moment.  Really need to set up a flow hood but probably not till spring when I can get some more room.


Edited by OysterFarmer, 17 February 2017 - 06:38 PM.


#6 dead_diver

dead_diver

    Stoned Since '76

  • OG VIP
  • 3,504 posts

Awards Bar:

Posted 17 February 2017 - 07:57 PM

I take spore prints on glass microscope slides. The slide can be put in a snack size Ziploc and when ready to make a syringe you just inject water into the bag, use the needle to scrape all the spores off the slide and suck the spore solution back up in the syringe.
Some things come sterile, or damn close to it, from the manufacturer. The inside of a new plastic ziploc bag before opening is sterile. As long as you practice good lab technique the success rate is excellent with this method. Just pay attention to the basics and you can't lose.
Needless to say the syringe will be dark, possibly black. If you aren't into spore overkill you can transfer it to a jar of sterile water and make more syringes.
This is a trick I use to fill syringes with sterile water. I buy a gallon jug of whatever water I'm using and I leave it closed. Do not open the jug! Next I wipe down a area just below the cap above and below the water line with alc. I then insert a needle through the plastic just above the water line and draw the water into the syringe. You can use small bottles of water but the gallon jugs have softer plastic. I use a large gauge needle to avoid getting it clogged with plastic when inserting it. Go to a feed store and tell them you want the biggest horse needles they have to fill printer cartridges. 21g will work but I prefer 18g or bigger. And with a spore solution that looks like mud a big needle helps to avoid clogs.
So now you have a syringe of sterile water and a Ziploc with a spore print on a glass microscope slide inside it. A finished spore syringe is about 30 seconds away.
You can take the prints on the slide using any method that works for you but I usually take the print inside the bag. That way I don't have to handle the slide or move it from one place to another in open air. I just tilt the bag while pinching a corner of the slide and drop the cap out of it. I don't zip it shut right away but instead leave it open for a while so any moisture from the cap can evaporate.
  • phlegmbae, MLBjammer, Heirloom and 2 others like this

#7 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 17 February 2017 - 08:16 PM

I take spore prints on glass microscope slides. The slide can be put in a snack size Ziploc and when ready to make a syringe you just inject water into the bag, use the needle to scrape all the spores off the slide and suck the spore solution back up in the syringe.
Some things come sterile, or damn close to it, from the manufacturer. The inside of a new plastic ziploc bag before opening is sterile. As long as you practice good lab technique the success rate is excellent with this method. Just pay attention to the basics and you can't lose.
Needless to say the syringe will be dark, possibly black. If you aren't into spore overkill you can transfer it to a jar of sterile water and make more syringes.
This is a trick I use to fill syringes with sterile water. I buy a gallon jug of whatever water I'm using and I leave it closed. Do not open the jug! Next I wipe down a area just below the cap above and below the water line with alc. I then insert a needle through the plastic just above the water line and draw the water into the syringe. You can use small bottles of water but the gallon jugs have softer plastic. I use a large gauge needle to avoid getting it clogged with plastic when inserting it. Go to a feed store and tell them you want the biggest horse needles they have to fill printer cartridges. 21g will work but I prefer 18g or bigger. And with a spore solution that looks like mud a big needle helps to avoid clogs.
So now you have a syringe of sterile water and a Ziploc with a spore print on a glass microscope slide inside it. A finished spore syringe is about 30 seconds away.
You can take the prints on the slide using any method that works for you but I usually take the print inside the bag. That way I don't have to handle the slide or move it from one place to another in open air. I just tilt the bag while pinching a corner of the slide and drop the cap out of it. I don't zip it shut right away but instead leave it open for a while so any moisture from the cap can evaporate.

I've already got lab quality giant syringes.  Also in a pinch a lot of people don't know that wal mart sometimes carries 'flavor injectors' which are basically large needles.  Not sure if they can pressure cook but they look like they could take it.

 

I've done the water from the jug too.  Usually though I just PC a quart jar with a home made lid with the gasket sealer.  Still is good to know all these different angles.  Never know when you might be in some end of the world scenario and humanity depends upon your ability to make mushrooms out of nothing.


  • MLBjammer and Heirloom like this

#8 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 17 February 2017 - 08:19 PM

I've been perusing the above mentioned archives.  Some won't load but that could be on my end.

 

I'm getting a better sense of it.  What I can't quite figure out though is how do you make sure the mushrooms ITSELF is sterile.  It seems like a mushroom growing in a dirty environment (like mine) will itself be dirty.  Or maybe I'm not understanding it.

 

Also getting the humidity off seems to be an issue.  That humidity is where a lot of my contams have come from I think.  Especially with wild harvested gourmets I've found.

 

Anyways I think I've got enough info to go at it again.  Still probably gonna order another syringe but that will probably be the last one.

 

Thanks for the help.



#9 dead_diver

dead_diver

    Stoned Since '76

  • OG VIP
  • 3,504 posts

Awards Bar:

Posted 17 February 2017 - 08:32 PM

That's the problem. The mushroom won't be sterile unless it's entire life cycle is carried out in a sterile environment.
I try to use caps that have just started to open to limit the gill and spore exposure to contams. I also wipe the cap with alc or h2o2 before placing in the Ziploc.
You won't get true sterility until you start doing the agar work from the prints.
A multi spore inoculation in a pf jar is usually clean but not absolutely sterile. It is a race between the many cube spores and the hopefully very few contams. The cubes usually win.
  • MLBjammer and Heirloom like this

#10 CatsAndBats

CatsAndBats

    [^._.^]ノ彡 & /|\( ;,;)/|\

  • OG VIP
  • 9,227 posts

Awards Bar:

Posted 17 February 2017 - 10:36 PM


There are mountains of information here at mycotopia already, I'd start here:

http://archives.myco...html?1061991123

Are we talking cubensis?
Are you trying to create a spore library or are you just trying not to spend more money? If you just want more inoculant purrhaps just 'milk' already colonized jars:
https://mycotopia.ne...ing-super-inoc/

Do you have an SAB/GB/flow hood in your employ? That will solve almost all of your dirty house problems, that and agar is your friend.

Trying not to keep spending money. Plus I'm sure they have some sort of record somewhere of all the times you order this shit. Once or twice they figure you are just some curious hippy. Do it again and again they probably look closer.

I just had a really good method. Order syringe. Put in a lc as soon as I open the box. Use that to innoculate grain. Went on like gangbusters but now trying to get the clean spores to work with just isn't going nearly as fast. Its like ten percent as fast as it was.

I've considered maybe injection some liquid into a petri plate as I use small mason jars with holes in the lids but my liquids haven't been doing as well.

Long term though I do want to not only have libraries but be able to walk in the woods and grab a mushroom then put it into culture. It would be impossible to farm the local mushrooms here by going through any distributor.

At what point did I suggest that one spend money?

#11 MLBjammer

MLBjammer

    Shaman Pouch!

  • Moderator
  • 9,419 posts

Donator


Awards Bar:

Posted 18 February 2017 - 07:11 AM

Maybe do a little Chronic style grow in a quart with a cake or something small and self-contained.  I find prints from invitro grows to be very clean.

 

I mean, you can take one print, inject that into a pint or quart of water, and you have dozens of syringes right there.  

 

Like Cat and DD said, a print is almost never even close to sterile.  It's just clean that you are shooting for.


  • CatsAndBats and Nonsanto like this

#12 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 18 February 2017 - 06:05 PM

That's the problem. The mushroom won't be sterile unless it's entire life cycle is carried out in a sterile environment.
I try to use caps that have just started to open to limit the gill and spore exposure to contams. I also wipe the cap with alc or h2o2 before placing in the Ziploc.
You won't get true sterility until you start doing the agar work from the prints.
A multi spore inoculation in a pf jar is usually clean but not absolutely sterile. It is a race between the many cube spores and the hopefully very few contams. The cubes usually win.

So the big companies then.  Do they grow them all in  totally clean rooms?  I've almost never seen a contam from my usual supplier.  I've heard a lot of the big companies all use the same few really big spore suppliers.  Often the big labs are in countries like Jamaica I think where there aren't laws against magics.  Or so I've heard.



#13 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 18 February 2017 - 06:18 PM

Maybe do a little Chronic style grow in a quart with a cake or something small and self-contained.  I find prints from invitro grows to be very clean.

 

I mean, you can take one print, inject that into a pint or quart of water, and you have dozens of syringes right there.  

 

Like Cat and DD said, a print is almost never even close to sterile.  It's just clean that you are shooting for.

Yeah that's probably what I should do.  I've even got half gallon jars I could do an entire grow in.



#14 CatsAndBats

CatsAndBats

    [^._.^]ノ彡 & /|\( ;,;)/|\

  • OG VIP
  • 9,227 posts

Awards Bar:

Posted 18 February 2017 - 07:23 PM

 

Maybe do a little Chronic style grow in a quart with a cake or something small and self-contained.  I find prints from invitro grows to be very clean.

 

I mean, you can take one print, inject that into a pint or quart of water, and you have dozens of syringes right there.  

 

Like Cat and DD said, a print is almost never even close to sterile.  It's just clean that you are shooting for.

Yeah that's probably what I should do.  I've even got half gallon jars I could do an entire grow in.

 

 

 

That sir is a brilliant idea.



#15 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 21 February 2017 - 06:12 PM

 

 

Maybe do a little Chronic style grow in a quart with a cake or something small and self-contained.  I find prints from invitro grows to be very clean.

 

I mean, you can take one print, inject that into a pint or quart of water, and you have dozens of syringes right there.  

 

Like Cat and DD said, a print is almost never even close to sterile.  It's just clean that you are shooting for.

Yeah that's probably what I should do.  I've even got half gallon jars I could do an entire grow in.

 

 

 

That sir is a brilliant idea.

 

Couldn't get my hand in the half gallons but this is what I came up with.  Described the soil mix here...https://mycotopia.ne...99813-new-tech/

Attached Thumbnails

  • SAM_6612.JPG


#16 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 21 February 2017 - 06:17 PM

This is what I came up with.  Put some tinfoil with holes on top.  Then put a cap on top in the glove box.

 

Seems like there are some spores in the water so we'll see in a few days if it germinates.  I've read though that contams can stick to the spores themselves though so we'll see.  The shrooms I've got are dirty as fuck.  The last plates I did are probably getting tossed entirely.  Insta green mold on all of them.  Live and learn I guess.

Attached Thumbnails

  • SAM_6591.JPG
  • SAM_6593.JPG
  • SAM_6592.JPG


#17 TVCasualty

TVCasualty

    Embrace Your Damage

  • OG VIP
  • 10,749 posts

Awards Bar:

Posted 21 February 2017 - 07:52 PM

Don't get hung up on "perfect" anything, for perfection is the enemy of excellence!


  • Sidestreet and CatsAndBats like this

#18 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 23 February 2017 - 06:04 PM

Nothen yet.  But also no dirty gunk.  I'll keep you guys posted.



#19 OysterFarmer

OysterFarmer

    Mycotopiate

  • Free Member
  • 660 posts

Awards Bar:

Posted 24 February 2017 - 04:42 PM

This is pretty ghetto but it seems to work.  I put some colonized grain into some LC in the glove box.  Took off pretty good after a few days.  The colonized myc floats which continues to impregnate the fluid apparently.  I'm just not having a lot of luck with the agar to grain part.  But if I put a big douche of innoculated water into a grain jar it seems to take off pretty good.  I've got enough grain atm at least to do a bunch of these.

Attached Thumbnails

  • SAM_6675.JPG


#20 Nonsanto

Nonsanto

    Mycophiliac

  • Free Member
  • 34 posts

Posted 10 October 2017 - 05:31 PM

Maybe do a little Chronic style grow in a quart with a cake or something small and self-contained.  I find prints from invitro grows to be very clean.
 
I mean, you can take one print, inject that into a pint or quart of water, and you have dozens of syringes right there.  
 
Like Cat and DD said, a print is almost never even close to sterile.  It's just clean that you are shooting for.


Yep yep yep. In "Growing Gourmet and Medicinal Mushrooms" (Stamets, p. 134), Paul talks about making "spore-mass masters".
Basically, just what you said MLBjammer. Fruit invitro on sterilized wood chips (for the example in the book) and aseptically removed, then sloshed around in a jar of sterilized H2O.
That would make spore syringes.
Or, just remove a nice looking fruit (or whatever you can get ahold of!) and remove (carefully) with sterilized tongs, tweezers, fork, spoon, wtf ever (in front of a laminar flowhood, in a glovebox, still-air box, etc). And then print.

Personally, I've not tried it yet, but sounds like it would be great for when you need absolutely clean, viable spores.
I use agar so I'm not real anal about how "clean" my prints are.
I can always transfer away from any Contams.

With that said, when I do print, I lay a big piece of aluminum foil down in front of the flowhood and place caps to be printed on that.
I got a bunch of Petri dishes, that once arrived, had a cpl holes in the plastic sleeves, I won't use them for culturing, but I will wipe them with isopropyl alcohol, let them dry in front of flowhood, then place one half of a Petri dish over each cap.
This keeps the humidity a little higher which helps the caps drop their spores.
After they drop their spores, I cut each print out of the foil and place into a sanitized (not sterile!) Petri dish, wrap with parafilm and store in the fridge.
Probably overkill, but whatever, I get bored. :-)




Like Mycotopia? Become a member today!