What qualifies as a sterile print?

[medicinedan]

So we have all read many teks on different sites about spore printing. I wont post a link now unless someone requests one.

So in theory i have mushrooms grown in a controlled enviroment,

they mature

I pick them with gloved hands and as carefully as possible set the caps down in a glovebox on some computer paper or foil that has been sprayed with 70% ethanol and allowed to dry in the glovebox

when the prints have dropped sufficiently (12-24hr something) put them in fresh baggies in the sterile glove box.

does this qualify as a 'sterile print'?

Could I truthfully claim to have 'sterile prints'?

In analysis, and technically speaking the prints may or may not be sterile. Yes most avenues were pursued in attempt to make the print as sterile as possiblem, within reason. A good percentage to 100% may in fact be sterile.

To be 100% sure of sterility though one would have to go to ridiculous measures such as growing the mushrooms invitro and then printing in a glovebox/ flowhood and printing onto autoclaved mediums and placed into gamma sterile bags.

I very much doubt that many people would grow completely invitro.

everytime you mist the enviroment or provide unfiltered air there is a good chance that contaminants may find their way onto the underside of the mushroom.

By no means am i discrediting peoples efforts at minimising contams. I just get confused. Is there an accepted, standard, sterlility level?

eg; If I print 100 in a dirty open air enviroment and later test 10prints for viability/ contams and find that out of the ten, all ten are viable and zero contams are present I could state statistically that 100% are sterile even though no sterile processes where adhered to.

What do the mycos think on this?

Cheerio

[gerbil]

The way I interpet it is more so 'clean print' rather than technically sterile.

Indoor fruiting, caps collect just as the veil breaks or let mature more and printing in a relatively closed environment is what i'd classify as clean.

IMO i don't think we need to make printing such a technical process such as heating foil etc. If there is contam, it gets left behind in the plate of intial spore streak and it finishes there IME.

I think a 'sterile print' is a little flawed coming from home cultivators, yet not unachievable, but I believe clean print would be a better option to use and customers shouldn't get so hung up about having 100% pure spore.

my.2c

[naja naja]

Wat I would concider "sterile" or "clean" is one that u could use straight in a LC or straight to grain with no contam, or if there is contam this is because of u, not the print.

Agar is only nessisary if u have a dirty print and want to clean it up. Like a print from the wild.

IME clean prints can be achived reletavely easily at home with low tch equipment. Even on paper and still be "clean"

I do think that "clean" is a much more appropriate term, because they are poss far from sterile.

Oh, and how technical is putting alfoil in the oven for 10-15 min? Put it in, sit on ur ass and smoke a bowl. Hey presto, clean alfoil.

And to see if u printing is "clean" streak a few plates with random print samples and c wat contams u get. Hopefully u will get none.

Edited July 3, 2006 by naja naja

[foolsbreath]

I believe removing the outer coating of the mushroom in a clean environment, and then allowing a light print develope before repositioning on a sterile plate with no agar in it or a piece of sterilized foil to drop the remainder of the spores gives a good clean print. I use a cupboard with a ozone generator in it running for several hours before making the print.

[occidentalis]

Personally I would only feel comfortable applying the term 'sterile' to a print prepared in a laminar flow from a mushroom grown in very clean indoor conditions.

Although, FWIW, sterile is not the correct term - in fact it indicates that there is nothing alive, including shroom spores. The word you want is 'axenic'.

[naja naja]

Personally I would only feel comfortable applying the term 'sterile' to a print prepared in a laminar flow from a mushroom grown in very clean indoor conditions.

Although, FWIW, sterile is not the correct term - in fact it indicates that there is nothing alive, including shroom spores. The word you want is 'axenic'.

Even grow the shrooms inside a laminare flow hood, that would be nice!

"STERILE" Hehe, I had a thought once before that u could rip a whole bunch of people off by selling truely sterile prints, where everything on it was dead. Tech correct, but I don't think the people would see it that way. But atleast the law prob would, since they r meant only for microscopy anyways, dead spores would stile be fine for Id I'm assuming.

I think I'll use the term "CLEAN" from now on.

[Rev]

yeah clean is better phrase to use

foil is already for practical purposes sterile when it comes off teh inside of the roll

better to bake it tyhan to sprayt with etoh

or better still to autoclave it for 15 mins at 15psi

To be 100% sure of sterility though one would have to go to ridiculous measures such as growing the mushrooms invitro and then printing in a glovebox/ flowhood and printing onto autoclaved mediums and placed into gamma sterile bags.

its not as hard as it sounds - really

i mean what you say is no more complicated than tissue culture of african violets

of course u need funds to buy all the equipment but after that u could train a technician from scratch to do it easily

[gerbil]

Oh, and how technical is putting alfoil in the oven for 10-15 min? Put it in, sit on ur ass and smoke a bowl. Hey presto, clean alfoil.

More so than placing directly on a sheet of paper...but I like the rest of your process ;) hehe sorry just playing. It definately depends on your desired goal. I prefer to grow on agar at this point in time, but can understand the need for a cleaner source if going for LC or syringe, but even then i'd personally prefer to agar it up, isolate and then put into LC or a syringe. But that's what I like about this field, you can achieve the same goals with the choice of so many paths.

Agar is only nessisary if u have a dirty print and want to clean it up. Like a print from the wild.

Not all the time, what if the desired goal is for specific isolates and not multispore innoculation?

lol sorry, just being picky take care mate.