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spores>Less = better? Thermal Death ? Hydrating ? [merged]


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#1 the_other_chap

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Posted 10 March 2005 - 05:48 PM

I love Shedthemonkey's method of laminating spore prints, but it seems to be damned hard to find cold laminating equipment & pouches in this neck of the woods.

I've been experimenting with a hot laminator, and I don't think the spores would have a problem withstanding the brief heating.
I guess the only way to be sure is to germinate some spores that have been heated, but I'd like some opinions from the floor to give me an idea of what to expect.

#2 Guest_yidakiman_*

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Posted 10 March 2005 - 09:20 PM

I'm fairly certain that thermal death occurs at 105F

#3 Guest_Peter Cottontail_*

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Posted 10 March 2005 - 09:22 PM

Pasteurization temperatures kill fungi spores. That's 140+ for an hour. I would think a brief exposure wouldn't hurt. You germ any of those comic prints yet shed?

#4 the_other_chap

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Posted 10 March 2005 - 11:43 PM

I'll give it a try tomorrow and see. I'll use two halves of the same print, hot laminate one, put the other in a baggie, prepare a syringe from both and see what happens. It's not going to be a particularly good test, as with the numbers of spores involved, even if a good portion are killed, I suspect there'll still be enough to get a vigorous germination going, and I don't know if I'll see much difference. I could try diluting to get less spores, but I don't have a US bath, so I can't be sure of even distribution and accurate results.
I think the laminator operates at around 100C but it's only applied for a couple of seconds as the sandwich passes through, and it cools very quickly once it's out.
I'll have to open up the laminator and see if both rollers are heated. If it's only one, I could turn the print to keep the Tyvek (and perhaps another insulating layer) between the spores and the heat, which might help.

I've germinated some of the "MonkeyTek" bookmarks (Tex and Z) and so far, all 12 jars (PF, six of each) are looking good. Healthy growth, and no contams. I'll post some pix when I fruit them. I'll probably use one of each to spawn rye grain to see how they do with casing.

Thanks for the info Roger, the prints have been in dessicant jars for a few days, so they should be bone dry and as tough as they're going to get :)

Yidakiman that sounds like maybe the temperature for thermal death of the mycellium?

#5 destroy_erase_improve

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Posted 10 March 2005 - 11:58 PM

i guess it also depends on how much space you allow also. if you have a small print maybe you want to give a half inch space at each side from the print itself. a little breathing room maybe.

#6 Guest_thafunkyone_*

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Posted 11 March 2005 - 02:57 AM

Someone germinated a print from a shed bookmark..redrum I think.

#7 the_other_chap

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Posted 11 March 2005 - 03:08 AM

Yep, no problems with Shed's, but he uses a cold laminator (EZ Laminator from Walmart as I understand). Unfortunately, they don't seem to be popular in the UK, so I've had to go with a cheap hot one for experimental purposes (£5 for an A6 laminator).
Chinagraph pencils seem to be an "order only" item now too. :rolleyes:
What do you guys use for writing on jars?

#8 Guest_Peter Cottontail_*

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Posted 11 March 2005 - 12:06 PM

"What do you guys use for writing on jars?"

'Sharpie' or other felt tip permanent ink marker.

#9 the_other_chap

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Posted 11 March 2005 - 04:28 PM

I'll have to look out for some.

Well, the experiment is underway :)
I innoculated two PF jars, one with the heated spores, and one with the control.
I hope it works, it was really easy to make a syringe from the laminated print, much easier than the bagged one.
If all goes well, I'll drag out my crap digicam and post a pictorial of the process. There's nothing new in it, it's just bits of other methods combined with shedthemonkey's "bookmark" prints.

#10 nomoreusmc

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Posted 11 March 2005 - 05:48 PM

how do you get the spores back out? isn't their a stick substance on the inside of the plastic? wouldn't that trap the spores

#11 the_other_chap

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Posted 11 March 2005 - 06:15 PM

You put the print in a little plastic pouch before it goes into the laminating pouch. 'Monkey uses acid free archival plastic, but I've just cut the corner off some baggies :)

If you leave a couple of millimeters each side of the print, there's enough room to snip the end off and either take the print out, or do what I did, and make a little cup out of it by squeezing the sides, and just squirt sterile water in and suck it out again a few times. (I hope the description makes sense, I'll try and post some pix if it proves workable, but my cam is poor & doesn't do closeups, so it might not be very spectacular.)
I think Shedthemonkey has posted a thread on his method somewhere, but I can't find it at the mo :(.
edit: here we go: http://mycotopia.net...read.php?t=2208

#12 the_other_chap

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Posted 15 March 2005 - 04:51 AM

Posted Image Good news everyone!

I've just checked my test jars, and the one from the laminated half of the print seems to be doing just as well as the unlaminated one, both seem to have good germination, and have good patches of mycelium starting to spread from the innoculation points.
So, within the limits of the experiment (which are considerable!) it seems that hot lamination doesn't damage the spores too much.
I don't know if there are any problems with the laminating plastic or the glue that might damage the spores' viability in the long term, but I think I'll be doing a lot of my prints this way in the future, so we'll see.

#13 chill

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Posted 13 June 2005 - 01:16 AM

Quote taken from another thread:

Another thing to consider, the more spores that germinate in your project, the worse it will do. That is a fact. You don't want so many sub strains that they are competing with each other. I cringe when I see very dark syringes, because I know that a very dark syringe is an almost guarantee of a poor harvest, while a totally clear to the naked eye syringe will almost always do very well.
RR



Whoa, now there's a provocative statement ;)

The trouble with a clear syringe is that you don't know how many spores are in it.

I've been having trouble getting good germination from 'crystal' vendor srynges. I was never sure if the jars didn't have good germination due to the jar itself or the syringe.

As a test I made up some black syringes from prints and had massive germination in the same, formally barren jars. I was also able to birth the 1/2 pint jars 12 days after innoculation. So more spores = better germination and colonization.

However, I'm fruiting the cakes right now and will post some pics of the harvest.

#14 panic

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Posted 07 October 2005 - 09:22 PM

this would have opened up a can of worms a while back....lol in fact, it did!
i think ali and rr debated over it for a while.

#15 I_am_me

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Posted 07 October 2005 - 09:50 PM

I've gotten used to making 5 prints from a syringe constantly. Nothing but complete success. I can see a point though where enough is enough and it just comes down to waste.

#16 Guest_freakachino_*

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Posted 07 October 2005 - 10:37 PM

my conclusion is based on print to syringe or liquid culture to substrate, no agar.
My results are inconclusive, fruits and yields roughly about the same either way. :)

#17 chill

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Posted 08 October 2005 - 04:02 PM

Whoa, did I create this thread and then forget about it?


Anyways, the cakes did just fine as far as yield went. No complaints. Can't remember the total weight but I was quite content.

I think yield was more influenced by strain than by spore volume. They were GT's if I remember correctly and they seem to be a great stain. Many of the other strains whose PF cakes were created using less spores produced lower yields.

#18 blackout

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Posted 09 October 2005 - 07:56 AM

if you suck up less spores, you also suck up less contams.

#19 chill

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Posted 09 October 2005 - 03:56 PM

if you suck up less spores, you also suck up less contams.


All one's prints should be clean. I think this is a separate issue.

My personal feeling is that until someone can prove that more spores = less yield then use more spores. They are basically free and reduce colonization time so why skimp?.

#20 Hippie3

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Posted 09 October 2005 - 04:13 PM

if you suck up less spores, you also suck up less contams.


this is relevant if there is concern about a dirty print,
and for agar work, primarily.
less relevant for clean prints and mass inoculations.




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