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LC syringe proof of concept


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#1 chill

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Posted 14 February 2006 - 06:21 PM

Hi all,

Bluehelix mentioned something called a vacutaner for storing liquid cultures.

That got me thinking that wouldn't it be possible to use steralized 4% sugar water to make spore syringes. That way the spores would germinate while they are in the syringe and would begin to immediately colonize the jar when injected. This would be helpful if you have a few minutes to make some syringes but plan on doing the jars later in the week for example.

So I tried this using maple syrup about a week ago and haven't noticed any growth in the syringe. I plan on innoculating wednesday and will see how it goes.

Has anyone else tried this and does anyone have any suggestions as to how to go about generating growth inside a syringe?

#2 Guest_freakachino_*

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Posted 14 February 2006 - 06:46 PM

Well, it sounds like a doable idea. I'd go with 20 cc or 60 cc syringes myself. That way you are creating a larger solution in the syringe.

So 4% of a 10cc syringe would mean, .25 cc/ml of karo or honey or vanilla. My math may be wrong ;)

I need to do a few syringes and have some Z spores to try it with.

And since my spores are already in water, I'm going to pc .25 cc of karo in the syringe, then suck up the spore water. I'll let ya know how it goes. :)
I'll put the syringes in ziplock and let em sit room temp.

#3 motograter

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Posted 14 February 2006 - 07:13 PM

I was thinking about this today too. Why not just make up a jar for LC the normal way, then take a syringe that you only have a small amount left in, then suck up the LC? I'm going to try it this way tomorrow. (All out of Vanilla....)
Freak, keep us posted on your attempts!
I'll do the same

#4 nepenthes_ak

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Posted 14 February 2006 - 07:20 PM

I was just thinking about this the other day... hmm yes please let us know.

Cheers

#5 chill

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Posted 14 February 2006 - 07:22 PM

My motivation for doing the LC in the syringe is because I have bad comtam issues with LC jars.
The syringe should be airtight and thus safe against contams.
'Freak, you may want to incubate the syringe rather than letting it sit at room temp. I think you've mentioned that your place is cold and cool temps would stop a syringe from otherwise germinating.
Cool, thanks for your input guys. I think this could really speed up the process.
I'd like to get the total time from innoculation to cropping down to a month if possible.

#6 anticheffy

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Posted 14 February 2006 - 07:32 PM

Im thinking the air tight thing would work against you.......

#7 chill

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Posted 14 February 2006 - 07:40 PM

I just checked my spore stash and I have a couple of prints left over.
They are GT and very nice fruiters.
I have a proposition for someone. I will donate a GT print to the first person to successfully create an 'invitro' LC syringe. The spores and water must be mixed together in the syringe and kept there until mycelium start to grow. Pics will be required.
I'll mail out the print once the tek has been verified by another board member. I'll also send out a print to that person (kind of like an assist in hockey) :)
I'm kind of low on spores or I'd offer more but I think this is a fun way to get things rollin'.

#8 chill

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Posted 14 February 2006 - 07:42 PM

Yea, that is a possibility. I've tried introducing an air bubble into the syringe as one way of dealing with it.

Also, I've seen mycelium in a syringe before and hip has commented on this before so I am hopeful. I believe it is possible.

Im thinking the air tight thing would work against you.......



#9 Guest_freakachino_*

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Posted 14 February 2006 - 08:01 PM

My tap water grows from spore water, so I'm thinking adding some karo in the mix should speed it up.

Chill, my room temp is 75 with my space heater so I figured room temp. Though maybe I'll throw some in the incubator too. See the time process at different temps.

This will be fun experimentation. Though I'm wondering if the mycelium syringe tek of shooting sterile water into a colonized jar and shaking then sucking that up works better. (I've done that method and its a good one).
Those syringes have to be used immediately though. The LC syringes could be stored.

#10 anticheffy

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Posted 14 February 2006 - 08:12 PM

theres a pic of a myc syringe somewhere here that has what appears to be a sclorotia grown out in it, it was so old.....I cant find it though

#11 chill

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Posted 14 February 2006 - 08:16 PM

Hi 'freak,
I forgot about that tek with the colonized jar. Thanks for the reminder.
'Course you have to have a jar sitting around to use and this way you could just have a print (like I have) :)
I think I'll try some potatoe water this weekend. I had OK results with this one time.

#12 chill

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Posted 14 February 2006 - 08:25 PM

I boiled my water to kill any contams. I was wondering if boiling removes O2 so I did a little research.

I found this little nugget on a gov't web site:
"The solubility of oxygen and other gases depends on water temperature. The colder the water, the more gases it can hold. Boiling water removes all gases."

Maybe reverse osmosis water would be better to use?

Also, shaking the water appears to put O2 back into it. Possibly having a bubble in the syringe and then shaking it would do the trick.

#13 BuckarooBanzai

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Posted 14 February 2006 - 08:40 PM

Perhaps leave a good bit of air (1/4 of it) and toss in a BB or two? That way you can shake the syringe to keep oxygen in the solution and break up the developing mycelia.

I believe air tight will work as long as you leave a good bit of air inside the syringe. The LC will need some air.

I totally agree with using a 60mL syringe, too.

#14 chill

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Posted 16 February 2006 - 05:36 PM

Have been thinking about the heat and air thing.

I decided to get some 'clean' water (reverse osmosis) and make a syringe out of it without heating it.

I bought a one litre bottle, dumped out half of it, shook it up real good to put some air in it (just in case).

Then I added 25ml of maple syrup and shook that up real good. I then pushed a syringe needle into the bottle and sucked up liquid to use in making spore syringes.

One syringe is a 30cc sucker that I have left 10cc of air in for shaking purposes. The other is a regular 10cc.

I've got two syringes in total down now for incubating. Hopefully this will show some results.

I used my first attempt from last week to innoculate some GT jars today. We'll see how they perform compared to the regular syringes I also used as side-by-side.

#15 motograter

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Posted 16 February 2006 - 05:45 PM

I just started 1 syringe, I had a little under .5cc of GT water, made up a LC jar, and sucked up 8cc of that. I made sure to leave enough room to shake, a little. now its in my inc. chamber. We'll see in a week or 2 if its working out.

#16 Guest_freakachino_*

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Posted 16 February 2006 - 06:36 PM

Here is what I did. My syringes allready have mycelium growing in under 24 hours.

What I did:

I had a jar of strain Z (from you chill :) )spore water. So in a small cup I (sorry didn't measure accurate lol) put about a teaspoon of karo, added 10 cc's of hot tap water. Stirred it around well. Sucked up 1 cc of the solution, and 1cc of air. Pc'd for 25 minutes at 15 psi.

Then I sucked up the spore water into each syringe and left that 1 cc of air.

On the last syringe, I shot the karo solution into the jar, then sucked up spore water. So now, even the jar is a lc along with 5 syringes of lc. I wasn't expecting this so quickly.

I'll get some pix later, also, I'll redo the procedure with photo's of all the steps. Pretty sweet idea really, think I'll inoculate some jars this weekend with it to see what happens :)

#17 chill

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Posted 17 February 2006 - 02:21 AM

Cool,

looks like we may have the first winner.

Pix and a mini tek write up would be sweet.

I'm going to refrain from checking on my syringes until at least 24 hours have passed.

#18 arezap

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Posted 17 February 2006 - 02:33 AM

Im thinking the air tight thing would work against you.......


Actually anticheffy, faof made some vLC's with 3 holes that were plugged with silicone caulk. faof pc'd the jars with toothpicks under the lids and allowed them to cool overnight to avoid cooling air in the jar creating a vaccuum. When opened the pc was opened the jars were sealed tightly. To load a syringe faof flames the needle and draws sterile air in the syringe, lets the needle cool, injects the sterile air into the jar, turns the jar upside down, and then loads the syringe. They have excellent results so far. The cultures have not stalled their growth and are shaken a few times daily.

#19 chill

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Posted 17 February 2006 - 02:40 AM

Just had a thought.

It was on my mind why people who pc their LC jars have success and yet boiling my water resulted in contams and no growth.

If the jar (or syringe in 'freaks case) are closed then maybe the air doesn't get boiled out entirely due to pressure but an open pan would dissipate all the O2.

#20 Guest_freakachino_*

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Posted 18 February 2006 - 01:45 PM

Here are some pictures of my syringes yesterday. I'm going to shoot them to some popcorn tonight or tomorrow to see how they do.

The spore water was pretty heavy, so that may have been a help.

Attached Thumbnails

  • ZKaroSyringes2.jpg
  • ZKaroSyringe1.jpg





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