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Signs of Successful and Unsuccessful LC


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#1 Flatbird

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Posted 21 October 2010 - 06:57 PM

What are signs of a contaminated or unsuccessful LC, as well as some of a successful LC?

#2 Nobody

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Posted 21 October 2010 - 09:30 PM

An LC that is bad shows off-color growths or cloudiness. If swirled in front of good lighting w/ some sort of dark background, it's easier to see this. A clean and viable LC should remain clear and have puffy or stringy tissue growth. That said, you can never be sure of the cleanliness of a culture until you test it on a jar or two.

#3 mydarling

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Posted 21 October 2010 - 09:33 PM

yeah, even an LC that "looks" clean could still be contaminated.

best way to know for sure is just do a few test jars with the LC before you do a whole batch of jars.
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#4 Earl Grey

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Posted 25 October 2010 - 07:56 AM

Maybe my latest PF Tec from a LC is of some help.
A little note upfront, i have, and i repeat, i have no intention to hijack your thread. I was going to post my results anyway and i thought i could do it here as it has the same question.
I had made a honey/water LC from a minuscule amount of B+ left over from a vial i ordered over the net.
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After a few days of swirling i could see some flakes, floating in the LC.
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After some more days i inoculate 4 PF Tec jars withe the LC.
In days i some points of the nasty green points showed, they quickly got overrun by the mycellium and after breaking 3 jars for study, i birthed the last jar with great succes. Yield was 11 grams dry. Not bad for a standard PF Tec jar.

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The initial contamination of TRICH was worrying me a little so filled a sterile syringe withe the LC as backup and injected 5 CC of 2H2O to the LC of 90 CC.
Last fry day i inoculate 5 PF TEC jars at different positions in the jars and left 1 jar for control.
1 Jar was inoculate close to the bottom, 1 halfway the jar and the last 1 just under the barrier. 1 Jar was excluded due to improper handling.
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Within days there was that nasty green stuff we all hate. And exactly where the fluid from the syringe has flowed over the glass.
No doubt that syringe was contaminated. Still the mycellium grows aggressive anyway.
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Back to your question, the syringe had no differences in color, cloud apparency or anything else. kept sterile at 2.4 C in my fridge for 6 weeks now.
Without the use of detailed info or a microscope, i can,t tell by just looking at it.
Now the good part..the LC itself, kept in the same conditions, showed dark spots on the inside of the glass. They appear on spots above the level of the jar and are most likely formed by swirling the LC.
I post some pics below in a minute.

I guess the only sure way to detect contamination is probably by using it and hope Gaya is on your side...
Hope this may help a little bit?

Attached Thumbnails

  • LC 002.jpg
  • LC 003.jpg

Edited by Earl Grey, 25 October 2010 - 08:19 AM.


#5 Flatbird

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Posted 25 October 2010 - 01:29 PM

It's all good, so the mycelium was flourishing beautifully and I was preparing to do some test jars when my buddy, instead of swirling the marble, shook the jar. Marble broke the jar and karo water went everywhere!! So I'm doing round two as we speak...

#6 Nobody

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Posted 25 October 2010 - 07:12 PM

Aw, that sucks! :eusa_booh But now you and him both know not to shake a jar w/ marbles! Sometimes you gotta find the good in the bad. Good luck w/ the second attempt.

#7 Freaky

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Posted 25 October 2010 - 07:19 PM

That sucks, broken karo lc's have happened to me before.

I quit using any marbles or broken glass or anything. I find it's just as good to use a suck and eject method when I'm ready to use the lc.

With your sterile syringe inserted into the jar you can suck up the mycelium and eject it forcefully back out over and over until it's broken up enough to make a final pull of broken up mycelium. It will also keep it macerated for the next set of syringes you're going to pull from the jar and use or store.

#8 demetrius shardson

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Posted 25 October 2010 - 08:11 PM

ouch! that blows man. I also stopped using anything in my LC's i just use a large enough jar to leave plenty of space inside the jar to break up the myc

#9 eastwood

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Posted 25 October 2010 - 08:45 PM

When making a lc its all in the lid of the jar/some sort of sterile air exchange..Two rtv self healing injection ports and a airport syringe is the way to go.Another big thing :amazed: is to only fill your container half full (lc solution) at the most.This will leave some room for the lc to breath in the jar.

#10 Nobody

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Posted 26 October 2010 - 09:39 PM

When making a lc its all in the lid of the jar/some sort of sterile air exchange..Two rtv self healing injection ports and a airport syringe is the way to go.Another big thing :amazed: is to only fill your container half full (lc solution) at the most.This will leave some room for the lc to breath in the jar.


Completely agree! I prefer LC jars w/ no filter media so that you can shake the livin' piss out of it to break it up. Filling half-way also gives the culture more air.

Freaky, couldn't the suck and shoot method be a possible invite for contamination? I guess you'd be pretty safe when working in a sterile glovebox or flowhood, but I just visualize a bad spore or two dropping down into the body of the syringe then getting into the solution when drawn up and shot again. Just a thought, if you have good luck w/ it then keep on keepin on! :headbang:

#11 Flatbird

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Posted 27 October 2010 - 10:18 AM

For shizzle...I Used one again but won't next. My present LC is hawian and looks good sitting in my incutator!!

#12 Freaky

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Posted 27 October 2010 - 10:33 AM

Completely agree! I prefer LC jars w/ no filter media so that you can shake the livin' piss out of it to break it up. Filling half-way also gives the culture more air.

Freaky, couldn't the suck and shoot method be a possible invite for contamination? I guess you'd be pretty safe when working in a sterile glovebox or flowhood, but I just visualize a bad spore or two dropping down into the body of the syringe then getting into the solution when drawn up and shot again. Just a thought, if you have good luck w/ it then keep on keepin on! :headbang:



No, and yes it's done in a glovebox or in front of a hood. You aren't drawing the string out at all. You insert it fully, suck it up, depress plunger fully, repeat as necessary, then make a final pull an withdraw the syringe.

#13 Jilkman

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Posted 27 October 2010 - 11:10 AM

Been doing my LC jars the same since the first I made. Only improvement I have made is less honey. Too much tends to leave clumps of honey material. Never really hurt, but just made the liquid look bad. Less honey = lighter color and easier to see contams. For my LC jars I use a single hole, and only half fill. I just shake the hell outta it occasinally. This keeps plenty of air dissolved, and keeps the mycelium broken up. I get lots of lil myco-puff-balls floating everywhere. Takes into the syringe real easy. No need for extra air exchange since this yeilds plenty to innoculate your substrates irreguardless.

Honey has worked well enough for me, I never even tried Karo. I use Wholesome Sweeteners - Organic Amber Honey, from the health food / organic section in the local grocer. Fill a pint jar about half full of water. Add about a teaspoon of honey. And just steam sterilize an hour in a fashion similar to BRF jars. No PC.

Just putting tape over the holes for now, but I think a self healing port would be optimal. Tho it would make it difficult to detect contams smell wise without cracking the lid open.

Other than the visable contams. You should be able detect them by smell if your not sealed too well. I have only had one LC jar contam. I first became aware by slowed growth in that particular jar. And confimed contam by smell. Just tossed it for simplicity.

As far as success... with the method I use, when I shake the jars I will see little myco-puff-balls floating around pretty easy when I shake the jars. Imagine each spore that germinates has 20+ tiny threads coming off of it in a 3-D snowflake like fashion. At least thats what I get. I consider them ready to use for innoculating substrates when I shake and its like one of those snow figurine jars, but about a quarter less snow. Settled the mycelium is about 20% of the jar, but it settles loosely.




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