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AyePlus
Stony Danza


Registered: 12/18/14
Posts: 3,393
Loc: Fairfield, Connecticut
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Using a microscope to inspect bulk substrate for contamination 3
#24873961 - 12/28/17 04:02 PM (6 years, 1 month ago) |
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When I’m looking at bulk substrates I occasionally see a different shade of mycelium in part of the tub, sometimes this is due to conditions and sometimes its a contaminant just starting to take ahold. The common reaction to this is to isolate the tub by taking it outside or at least into a different room and waiting to see if that snowy white patch was just wierd cube myc reacting to conditions, a minor bacterial infection, or if its going to be the mean green. I’m an impatient person by nature and this drives me crazy, I’d rather toss tubs that are fucked asap and the outdoor conditions where I live are unsuitable for putting tubs outside.
Instead of all that wait and see shit I take a sample and mount it on a slide so I can check it out under my microscope. Figured I’d share how I do this with y’all even though its mad simple.
Any biological or soil specimen microscope with 400x total magnification will work, the cheaper more popular ones are made by AMscope and Omax. Heres mine
 I bought Omax because I heard that since they are a Canadian company they use swiss optics which are better than anything else available in tha price range.(USA has higher tariffs on swiss goods than Canada. So any scope made here from same parts would cost more)
Once you have your scope set up with clean slides and cover slips ready its time to prepare your specimen, open the questionable tub and carefully using a (clean) scalpel remove a small amount of the target mycelium.


Place the smallest piece you can on the slide and put a single drop of distilled water on it

Then using your cover slip try to spread out the mycelium as much as possible.(its not much)

Ready to go.
Place the slide onto your microscope, turn the light on and center your sample in the light, start at the lowest possible magnification and once image is in focus move up from there.
Heres what it looks like at 100x

Cant really see much lets go to 400x
 a whole mess of mycelium, a few bacteria floating around and the cocofiber its all growing on, I dont see any aspergillis or trichoderma sporophores formed or forming and all the mycelium looks to be the same color and diameter, meaning its probably one species of fungi producing the mycelium. Some bacilli (rod shaped) bacteria are not ideal but in low numbers. Spawn was probably slightly bacterial, but since it was open air spawned you can’t be sure.
Make sure you check out the whole sample to be sure. I’m not seeing anything suspect, the bacteria are free moving in the solution, not in huge numbers and not clustered up on the mycelium eating it, the mycelium appears to have all of its cell walls intact, back onto the fruiting shelf, I’m glad too because it was beginning to pin and I really wanted to see this clones performance.
If it was trichoderma it would look something like this
 This is where that sample was taken from, to an untrained eye it could easily be misidentified it as knots.
 This one went straight to the compost pile, no hesitation, all those little round things on the end of the hypha are trich spores!!
And if it were aspergillis it would look something like this(not my image)

Unknown contaminant that appeared after cold storage.

Notice the thin wispy mycelia and the regular psilocybe mycelia intertwined.

Tubs that were spawned with grain inoculated with this agar were slow to fruit and the first flush pins all aborted or grew a white fuzz and were very small, with higher FAE the second flush did better but still opened caps prematurely, and the yeild was crap.
This post will be updated with more microscope photos as I take them. I will try to eventually include shots of a really bad bacillus infection, pin mold, cobweb and mycogone. I don’t plant on purposely introducing them into any bulk subs so they may be samples from petris instead.
. All photos taken with an phone 7 and this thing.

(Wasn’t sure if this belongs in Advanced or contamination or mush cult, mods feel free to move it, just figured cult gets the most use)
Edited by AyePlus (03/01/18 01:59 PM)
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Just_A_Noob
Breathing



Registered: 12/30/16
Posts: 6,809
Loc: PNW
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Re: Using a microscope to inspect bulk substrate for contamination [Re: AyePlus]
#24873976 - 12/28/17 04:07 PM (6 years, 1 month ago) |
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Cool
-------------------- Wearing a mask is bad for my physical, emotional, and spiritual health. Complying = Consent Wide Mouth 1/2 Pint No-Pour TEK TC Teks & Links
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Shrooms4life
TrippyShaman



Registered: 06/08/17
Posts: 1,375
Loc: Under the Viel
Last seen: 5 years, 8 months
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Just_A_Noob]
#24874035 - 12/28/17 04:28 PM (6 years, 1 month ago) |
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Awesome post man, does cutting the small sample affect the bulk sub in anyway?
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Icezizim



Registered: 11/09/09
Posts: 132
Loc: Oklahoma
Last seen: 10 months, 9 hours
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Just_A_Noob]
#24874069 - 12/28/17 04:38 PM (6 years, 1 month ago) |
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mushboy
modboy



Registered: 04/24/05
Posts: 32,464
Loc: where?
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Icezizim]
#24874085 - 12/28/17 04:42 PM (6 years, 1 month ago) |
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Quote:

thats trich no doubt. who said that was knots?
anyway cool idea. looks fun. think like a chef tho.. they dont take the foods temp to know when its done. they just know. same concept. but it could help with just visually disgusting good v bad when you are unsure.
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AyePlus
Stony Danza


Registered: 12/18/14
Posts: 3,393
Loc: Fairfield, Connecticut
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Re: Using a microscope to inspect bulk substrate for contamination [Re: mushboy]
#24874165 - 12/28/17 05:07 PM (6 years, 1 month ago) |
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Quote:
Shrooms4life said: Awesome post man, does cutting the small sample affect the bulk sub in anyway?
NoQuote:
mushboy said:
Quote:

thats trich no doubt. who said that was knots?
anyway cool idea. looks fun. think like a chef tho.. they dont take the foods temp to know when its done. they just know. same concept. but it could help with just visually disgusting good v bad when you are unsure.

Looking back at the old thread only one person guessed knots.
I am a chef and have worked as such and I occasionally use a thermometer just to make sure. Not saying you need to go out and buy a scope if you don’t have one, just suggesting a way to use one for double checking if you do have it or want to be extra sure.
I just threw out a box that had 1.5 qts in it because it started to go green and since one of the jars was split between that and this one I was being extra vigilant. When I saw the bright white spot on this one last night I almost tossed it but I’m glad I didn’t since it looks fine under the scope and is starting to pin.
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AyePlus
Stony Danza



Registered: 12/18/14
Posts: 3,393
Loc: Fairfield, Connecticut
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Re: Using a microscope to inspect bulk substrate for contamination [Re: AyePlus]
#25003990 - 02/18/18 03:51 PM (6 years, 5 days ago) |
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OP updated with new pictures of an unknown contaminant.
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Psilosopherr
A psilly goose



Registered: 02/15/12
Posts: 12,278
Last seen: 2 months, 4 days
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Re: Using a microscope to inspect bulk substrate for contamination [Re: AyePlus]
#25004099 - 02/18/18 05:03 PM (6 years, 5 days ago) |
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Cool idea Got a scope sitting right here that I never make use of
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Tormato  
The Goddess Kali Meh 😛




Registered: 07/01/17
Posts: 6,067
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Psilosopherr]
#25007908 - 02/20/18 12:07 PM (6 years, 3 days ago) |
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Quote:
Psilosopherr said: Cool idea Got a scope sitting right here that I never make use of
Glad I stumbled across this gem of a thread. I got a scope myself just collecting dust...well not anymore
-------------------- Helpful Threads The Shroomery Store Tormato's Q&A Thread Post Questions Here or PM me! "Lately it occurs to me what a long, strange trip it's been." ~ Grateful Dead Before you start...Do you have a Pressure Cooker and a Dehydrator? I highly recommend getting both!
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Germs
Space Force


Registered: 06/26/11
Posts: 4,607
Loc: Texas
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Tormato]
#25007922 - 02/20/18 12:14 PM (6 years, 3 days ago) |
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Awesome post, and nice scope!
--------------------
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van hatton
Still a noob



Registered: 11/23/14
Posts: 5,617
Loc: Michigan
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Tormato]
#25007935 - 02/20/18 12:19 PM (6 years, 3 days ago) |
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-------------------- If I ever give out misinformation please inform me so I can have the correct information. Tmethyl said: Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy. Caps McGee said:
Fun part is figuring out what works best for you
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Psilosopherr
A psilly goose



Registered: 02/15/12
Posts: 12,278
Last seen: 2 months, 4 days
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Tormato]
#25007977 - 02/20/18 12:36 PM (6 years, 3 days ago) |
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Quote:
Tormato said:
Quote:
Psilosopherr said: Cool idea Got a scope sitting right here that I never make use of
Glad I stumbled across this gem of a thread. I got a scope myself just collecting dust...well not anymore 
I'd been under the impression I needed fancy scheduled reagents to make use of a microscope for this hobby.
Then comes this guy who actually remembers middleschool science class just putting a sample on distilled water, making me feel quite the fool
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Tormato  
The Goddess Kali Meh 😛




Registered: 07/01/17
Posts: 6,067
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Psilosopherr]
#25008064 - 02/20/18 01:06 PM (6 years, 3 days ago) |
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Quote:
Psilosopherr said:
Quote:
Tormato said:
Quote:
Psilosopherr said: Cool idea Got a scope sitting right here that I never make use of
Glad I stumbled across this gem of a thread. I got a scope myself just collecting dust...well not anymore 
I'd been under the impression I needed fancy scheduled reagents to make use of a microscope for this hobby.
Then comes this guy who actually remembers middleschool science class just putting a sample on distilled water, making me feel quite the fool 
My thoughts exactly!
-------------------- Helpful Threads The Shroomery Store Tormato's Q&A Thread Post Questions Here or PM me! "Lately it occurs to me what a long, strange trip it's been." ~ Grateful Dead Before you start...Do you have a Pressure Cooker and a Dehydrator? I highly recommend getting both!
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verum subsequentis
seeker of truth



Registered: 03/22/16
Posts: 8,732
Last seen: 1 year, 8 months
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Tormato]
#25031377 - 03/01/18 10:28 AM (5 years, 11 months ago) |
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Mad Season
hookers and blackjack



Registered: 09/16/12
Posts: 12,666
Loc: Canada
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Re: Using a microscope to inspect bulk substrate for contamination [Re: verum subsequentis] 3
#25031520 - 03/01/18 11:36 AM (5 years, 11 months ago) |
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Another cool thing you can do is get a USB microscope or maybe even a pocket microscope, and you can potentially see through spawn jars at a contaminant. Pretty cool stuff!
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HamHead
Hard Ass Motherfucker



Registered: 03/17/15
Posts: 6,107
Loc: Galactic sector ZZ9 Plura...
Last seen: 2 years, 9 months
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Re: Using a microscope to inspect bulk substrate for contamination [Re: Icezizim]
#25031590 - 03/01/18 12:07 PM (5 years, 11 months ago) |
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Quote:
Icezizim said:

-------------------- The Italian researchers’ findings, published by the INT’s scientific magazine Tumori Journal, show 11.6% of 959 healthy volunteers enrolled in a lung cancer screening trial between September 2019 and March 2020 had developed coronavirus antibodies well before February. https://www.reuters.com/article/us-health-coronavirus-italy-timing-idUSKBN27V0KF This online first version has been peer-reviewed, accepted and edited, but not formatted and finalized with corrections from authors and proofreaders https://www.icandecide.org/
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