|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
ruckusman
Figment of my own imagination

Registered: 05/08/10
Posts: 190
Last seen: 4 years, 3 months
|
I might even be clever...
#13030981 - 08/10/10 09:58 AM (14 years, 5 months ago) |
|
|
So I have some LC's going in baby food jars approx 250 ml jars - half full, they've got self healing RTV silicone injection ports, but no GE holes.
Made that way deliberately so that I could shake them vigorously to speed growth, but I knew they would stall, which they have.
What I wanted was a way to inject some sterile air whilst loosening the lid a fraction to let the old air out.
How to get sterile air?
I did it like this...
Things you'll need
PC with the ability to remove the jiggler weight Large syringe with needle (gauge isn't so important) Alcohol lamp (for convenience) or a lighter Some alcohol swabs
Filled the pressure cooker with approx 5 litres of water and give it a good whirl for 15-20 minutes, that way all the air and water inside is sterile.
Let it cool till the jiggler weight can be removed without scalding myself, but I was wearing thick leather work gloves to be sure.
Then I've got a 60ml syringe with a 50mm 19 gauge syringe attached (previously sterilised) in a PC run.
I prepped each jar before starting by checking that I could turn the lid with one hand. They can get really tight and take hands two sometimes.
Crank the heat to the PC back up and wait till the steam starts to blow out the hole vigorously and then insert the needle all the ay into the steam vent.
Mine is good in that it's got a hole straight through so I can put the needle all the way down and use the plastic to plug the vent hole. Let the steam drive the plunger all the way up and pull it out.
Then straight out of the PC vent hole into the alcohol lamp flame till red hot and straight into the RTV injection port. I managed to swab the port with one hand whilst heating the needle.
Drive down on the plunger till it springs back then push it all the way, loosen the lid very gently till it doesn't spring back and then re-tighten.
I did mine twice so that they at least got 120mls of fresh steamed air.
The quicker that you go from the vent port to the alcohol lamp to the RTV injection port the less chance there is of the steam condensing inside the syringe drawing potentially contaminated air.
We'll see if my LC's speed up again in a few days, or maybe I just contaminated them all
I'm hoping for the best
Would love any feedback if anyone can think of any flaws in my reasoning or process
peace out
Edited by ruckusman (08/10/10 10:01 AM)
|
Issius
Stranger



Registered: 06/29/10
Posts: 510
Loc: Pennsylvania
Last seen: 10 years, 6 months
|
Re: I might even be clever... [Re: ruckusman]
#13031083 - 08/10/10 10:41 AM (14 years, 5 months ago) |
|
|
Well....it seems like it might work.
Generally when people need "sterile" air they just hole the tip of the syringe over a flame and pull air in from there. This is good enough to kill most things, but you could run into some problems.
A word of advice though, its a lot easier to use two layers of tyvek siliconed to the jar lid (one on top of the lid, one on the underside of the lid) so that you can shake it without the top layer getting wet. The wet filter "wicks" in contaminants, so if the top one isn't wet there's no issue. With that in mind, I have an LC that I've been shaking like crazy before use and only has one layer of tyvek. Maybe I've just been lucky, but no contams or problems from that one. (The original plan was to only use the LC once, which is why I shook).
I'm interested in seeing how this goes, but I think its a little more work than necessary.
|
FoxFire
Energized Matter



Registered: 07/17/05
Posts: 1,281
Loc: PNW
|
Re: I might even be clever... [Re: ruckusman]
#13031101 - 08/10/10 10:46 AM (14 years, 5 months ago) |
|
|
Personally Id just open the jar in a glovebox.
-------------------- Unless someone like you cares a whole awful lot,
Nothing is going to get better. It's not.
|
Issius
Stranger



Registered: 06/29/10
Posts: 510
Loc: Pennsylvania
Last seen: 10 years, 6 months
|
Re: I might even be clever... [Re: FoxFire]
#13031126 - 08/10/10 10:52 AM (14 years, 5 months ago) |
|
|
Quote:
ManaShaman said: Personally Id just open the jar in a glovebox. 
|
ruckusman
Figment of my own imagination

Registered: 05/08/10
Posts: 190
Last seen: 4 years, 3 months
|
Re: I might even be clever... [Re: FoxFire]
#13032938 - 08/10/10 05:41 PM (14 years, 5 months ago) |
|
|
Quote:
ManaShaman said: Personally Id just open the jar in a glovebox. 
AWW that's just mean Agreed though.
But I have minimised the possibility of contamination as I see it.
Plus drawing the air from within a flame will have both C0 and CO2 from the combustion
It sounds like a lot of work, but actually took all of 10-15 minutes to do 10 jars once the water was PC'ed and then boiling again
|
Base Icks



Registered: 03/19/10
Posts: 6,191
Loc: Shroomshire
|
Re: I might even be clever... [Re: ruckusman]
#13033009 - 08/10/10 05:57 PM (14 years, 5 months ago) |
|
|
If your pc was going for 10-15 minutes I bet most of the air was gone and it was just steam(H2O)
|
Breakfast Crew
Cheerios



Registered: 01/20/09
Posts: 1,985
Last seen: 13 years, 5 months
|
Re: I might even be clever... [Re: Base Icks]
#13033423 - 08/10/10 07:40 PM (14 years, 5 months ago) |
|
|
I tried PCing my water loaded syringes and the plunger blew out. Esta no bueno.
|
hamloaf
Q-dood ®©™√


Registered: 12/23/09
Posts: 24,389
Loc: ation: Based.
Last seen: 1 day, 2 hours
|
|
Quote:
So I have some LC's going in baby food jars approx 250 ml jars - half full, they've got self healing RTV silicone injection ports, but no GE holes.
Made that way deliberately so that I could shake them vigorously to speed growth, but I knew they would stall, which they have.
What I wanted was a way to inject some sterile air whilst loosening the lid a fraction to let the old air out.
You gotta' have that GE, my freind. This sounds like a case for "hippyflipping". Ever heard of it? Are you working with a glove box or a flowhood? Sterile air syringes are achieved by drawing in the air right over a flame (as mentioned earlier). HL
|
choempi
Stranger



Registered: 02/21/10
Posts: 52
Last seen: 13 years, 1 month
|
Re: I might even be clever... [Re: hamloaf]
#13033745 - 08/10/10 08:41 PM (14 years, 5 months ago) |
|
|
stick a filter syringe in, done.
|
hamloaf
Q-dood ®©™√


Registered: 12/23/09
Posts: 24,389
Loc: ation: Based.
Last seen: 1 day, 2 hours
|
Re: I might even be clever... [Re: choempi]
#13033760 - 08/10/10 08:45 PM (14 years, 5 months ago) |
|
|
Quote:
choempi said: stick a filter syringe syringe filter in, done.
There ya' go, big guy.  HL
|
choempi
Stranger



Registered: 02/21/10
Posts: 52
Last seen: 13 years, 1 month
|
Re: I might even be clever... [Re: hamloaf]
#13033843 - 08/10/10 09:00 PM (14 years, 5 months ago) |
|
|
Quote:
hamloaf said:
Quote:
choempi said: stick a filter syringe syringe filter in, done.
There ya' go, big guy.  HL

semantics
|
ruckusman
Figment of my own imagination

Registered: 05/08/10
Posts: 190
Last seen: 4 years, 3 months
|
Re: I might even be clever... [Re: choempi]
#13034179 - 08/10/10 10:05 PM (14 years, 5 months ago) |
|
|
Yep the syringe filters are now on my list
But I do like my improvisation all the same
Plus I do have an idea in mind for replaceable twin stage Tyvek filters that can mimic syringe filters
Will write a TEK for that once I've made my first - hopefully this evening
Sometimes it's about the workarounds and tricks with this stuff as it keeps it cheap and accessible to everyone
|
FoxFire
Energized Matter



Registered: 07/17/05
Posts: 1,281
Loc: PNW
|
Re: I might even be clever... [Re: ruckusman]
#13035108 - 08/11/10 03:13 AM (14 years, 5 months ago) |
|
|
Quote:
ruckusman said:
Quote:
ManaShaman said: Personally Id just open the jar in a glovebox. 
AWW that's just mean Agreed though.
But I have minimised the possibility of contamination as I see it.
Plus drawing the air from within a flame will have both C0 and CO2 from the combustion
It sounds like a lot of work, but actually took all of 10-15 minutes to do 10 jars once the water was PC'ed and then boiling again
Not trying to be mean, or funny, tho it happens sometimes. Seems like a lot of rigmarole for a simple task. Them more you dick around with your sterility the less sterile it becomes. KISS- keep it simple stupid. 
Oh, and I may be completely wrong here, but if you suck in air from over a flame, wouldn't there be no oxygen seeing as fire uses oxygen for fuel? The mycelium needs oxygen to grow big and strong right? I dunno, it all seems silly to me.  Good luck tho!
-------------------- Unless someone like you cares a whole awful lot,
Nothing is going to get better. It's not.
|
ruckusman
Figment of my own imagination

Registered: 05/08/10
Posts: 190
Last seen: 4 years, 3 months
|
Re: I might even be clever... [Re: FoxFire]
#13035283 - 08/11/10 05:40 AM (14 years, 5 months ago) |
|
|
Quote:
ManaShaman said:
Oh, and I may be completely wrong here, but if you suck in air from over a flame, wouldn't there be no oxygen seeing as fire uses oxygen for fuel? The mycelium needs oxygen to grow big and strong right? I dunno, it all seems silly to me.  Good luck tho!
I agree with this theory of the O2 being used for combustion which is why I used the steam to fill the syringe, the flame was to sterilise the tip of the needle
|
Base Icks



Registered: 03/19/10
Posts: 6,191
Loc: Shroomshire
|
Re: I might even be clever... [Re: ruckusman]
#13035331 - 08/11/10 06:44 AM (14 years, 5 months ago) |
|
|
I bet you still got all steam out of the pc and still got no O2.
Flame the tip of the needles till very red hot.
Then suck are in.
Easy peasy.
The flames out and the air gets super heated as it enters the syringe.
With the flame out you dont have to worry about not getting O2.
|
|