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Takeatrip
Stranger
Registered: 09/10/23
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SAB contamination, how? 1
#28647193 - 02/05/24 04:56 AM (3 months, 10 days ago) |
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Heh folks, I have a home made an SAB which I am very pleased with but recently I did a transfer from a BRF cake to agar into three petri dishes and so far one is contaminated. I am annoyed as I could not understand how it got contaminated. I cant remember the thread I posted my pics in but some suggested taking the tape off of the hand holes so it does not create a piston effect when pulling my hands in and out. Should I do this? I will post a pic or two below so you can see what I mean. Also does it matter if your forearms are not covered while working in the SAB? Do any of you use long gloves? And just to note I do try to work at the back of the SAB so my hands or arms are not hovering over an open dish while transferring. I have considered making a little stainless steel raised platform to do more work on and to see if it helps reduce any mould particles floating about in the SAB when I first go in. When I did the transfer, the BRF cake jar was clean and wiped down and in the SAB at the back and I carefully turned it over into a plastic container that was wiped down with a very mild solution of bleach followed my isopropyl alcohol. I used a flame sterilised x-acto knife (flamed outside of SAB due to isopropyl alcohol not long ago had been used in there) that is just for mycology and took some scrapings for the dishes. I did not have each dish open for long at all, maybe 20/30 seconds or so and they were lifted up by my other hand so they were now low down. Any suggestions on improving my SAB or technique would be great.
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DERRAYLD
Constructus


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Re: SAB contamination, how? [Re: Takeatrip] 2
#28647197 - 02/05/24 05:15 AM (3 months, 10 days ago) |
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Remove the covers on your holes, that defeats the fundamental of a sab. You're creating a gasket that is pumping air in as you move.
You really do need a raised work area because everything settles at the bottom of the SAB.
Edited by DERRAYLD (02/05/24 05:22 AM)
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JungleSkunk
Hashashin


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Re: SAB contamination, how? [Re: DERRAYLD]
#28647217 - 02/05/24 05:42 AM (3 months, 10 days ago) |
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The area you are working in doesnt look very clean. Probably the cause.
-------------------- ~I wanna grow up in truck stops and on friends floors... Maybe then I can feel my heart beat in rhythm with the real me.~
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DERRAYLD
Constructus


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Re: SAB contamination, how? [Re: JungleSkunk] 3
#28647219 - 02/05/24 05:44 AM (3 months, 10 days ago) |
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Nah, SABs can be used in dirty areas as long as the internal space is allowed to settle Nd you keep movement slow.
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AlexandrDughin
Explorer

Registered: 10/30/23
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Re: SAB contamination, how? [Re: Takeatrip] 2
#28647274 - 02/05/24 07:13 AM (3 months, 10 days ago) |
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The purpose of a SAB is to highly increase your success rate, but it will never be 100% success rate.
I get contaminations on my plates at a rate .. idk 2/10 plates ( 20% ) anyway.
steps you can do, as others said :
- remove the tape on the holes while working small holes = more air acceleration = more current = more contamination
- place a wet paper towel on the bottom, it will help keep the dust "locked" on the bottom.
- place a small box or something, to raise your working area, so the tools and dishes are away from the contaminated bottom.
- slow movements.
...
aaaand you will still get contaminated plates every now and then
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Takeatrip
Stranger
Registered: 09/10/23
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Quote:
JungleSkunk said: The area you are working in doesnt look very clean. Probably the cause.
My work area is in my loft, the air doesn't move much up there but it is the only place I have the space to do it undisturbed.
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Takeatrip
Stranger
Registered: 09/10/23
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Quote:
AlexandrDughin said: The purpose of a SAB is to highly increase your success rate, but it will never be 100% success rate.
I get contaminations on my plates at a rate .. idk 2/10 plates ( 20% ) anyway.
steps you can do, as others said :
- remove the tape on the holes while working small holes = more air acceleration = more current = more contamination
- place a wet paper towel on the bottom, it will help keep the dust "locked" on the bottom.
- place a small box or something, to raise your working area, so the tools and dishes are away from the contaminated bottom.
- slow movements.
...
aaaand you will still get contaminated plates every now and then 
Thanks. Appreciate your comment, I have taken notes and will try these out 👍🏻
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Takeatrip
Stranger
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Re: SAB contamination, how? [Re: DERRAYLD] 1
#28647417 - 02/05/24 09:42 AM (3 months, 10 days ago) |
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Quote:
DERRAYLD said: Remove the covers on your holes, that defeats the fundamental of a sab. You're creating a gasket that is pumping air in as you move.
You really do need a raised work area because everything settles at the bottom of the SAB.
I will remove them today. As for raising it up I will make something more permanent next week for it. Space is an issue, but it wont stop me.
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morningWood
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Re: SAB contamination, how? [Re: Takeatrip] 1
#28647439 - 02/05/24 10:19 AM (3 months, 10 days ago) |
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One piece of advice that really helped me was to work in the back of the box.
-------------------- 🅃🄴🄰🄼 🄽🄴🅂🄲🄾 You can lead a man to knowledge, but you can't make him think.
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Skropi
Space Ritual


Registered: 11/01/23
Posts: 414
Loc: Greece
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How many times have you worked in a SAB? I am asking because my first times I couldn't discern what went wrong, but after some time, I could easily tell what my mistakes were, and my success rate went way up, even when I am a bit sloppy. By the way, it slipped the attention of everyone, but you said that you didn't flame sterilise your scalpel inside the SAB because you have sprayed iso beforehand. You never use a flame in the SAB, as it will just create turbulence. Spray the inside of the SAB with some diluted soapy water, no need for bleach or iso, but do spray all the surface inside, wait for at least half an hour for everything to settle, and don't keep your hands and tools above anything that you don't want contaminated. Oh, I also think that your SAB is too small. I believe that size matters, bigger SAB, less turbulence. Get a nice, big, tub, mine is a 32lt one, and it is small, I am planning to make a custom SAB, with plexiglass.
-------------------- Ἄνδρα μοι ἔννεπε, Μοῦσα, πολύτροπον
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Takeatrip
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Re: SAB contamination, how? [Re: Skropi]
#28647575 - 02/05/24 12:21 PM (3 months, 10 days ago) |
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Quote:
Skropi said: How many times have you worked in a SAB? I am asking because my first times I couldn't discern what went wrong, but after some time, I could easily tell what my mistakes were, and my success rate went way up, even when I am a bit sloppy. By the way, it slipped the attention of everyone, but you said that you didn't flame sterilise your scalpel inside the SAB because you have sprayed iso beforehand. You never use a flame in the SAB, as it will just create turbulence. Spray the inside of the SAB with some diluted soapy water, no need for bleach or iso, but do spray all the surface inside, wait for at least half an hour for everything to settle, and don't keep your hands and tools above anything that you don't want contaminated. Oh, I also think that your SAB is too small. I believe that size matters, bigger SAB, less turbulence. Get a nice, big, tub, mine is a 32lt one, and it is small, I am planning to make a custom SAB, with plexiglass.
I have used my SAB for a few months now with some success, I have managed to get 2 strains of clean cultures so far. My tub is 54 litres which I believe is enough for the work I do and should be big enough for less turbulence but I will keep the idea in mind. I will be working much slower in the SAB from now on. As for flame sterilising the needle, I have always done it just outside the SAB as I don’t want any accidents. That plexiglass idea sounds good! Send me some pics when your done as I might try making one this year when the summer gets here, I will start having a look at dimensions etc.
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LadysKnight
Hello Ladies


Registered: 10/09/15
Posts: 1,976
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Re: SAB contamination, how? [Re: Takeatrip] 3
#28647625 - 02/05/24 01:04 PM (3 months, 10 days ago) |
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As said, your sab looks a bit small. If you think of air as water, the smaller your vessel, the more stirring your movements will make. The larger, the less impact. My diy sab is 4ft wide, 2ft tall, 1.5ft deep or 340 liters.
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Takeatrip
Stranger
Registered: 09/10/23
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Last seen: 10 hours, 31 minutes
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Quote:
LadysKnight said: As said, your sab looks a bit small. If you think of air as water, the smaller your vessel, the more stirring your movements will make. The larger, the less impact. My diy sab is 4ft wide, 2ft tall, 1.5ft deep or 340 liters.
I will see what happens with this one first now I have removed the tape and the air has settled. I have some more work to do in there so we will see.
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LogicaL Chaos
Ascension Energy & Alien UFOs




Registered: 05/12/07
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Re: SAB contamination, how? M.G.B.D.C. - an experimental suggestion [Re: Takeatrip]
#28648351 - 02/06/24 01:59 AM (3 months, 9 days ago) |
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So I am actually having a similar problem and I have a similar-sized SAB. I think that for SABs, they have to kinda big for them to work properly.
However, I have an idea. I want to bring back the glovebox, but only for small boxes not suitable as a SAB.
This is experimental but i think its pretty fesiable. Convert a glovebox from a too-small sized SAB. About a month ago, i experimented with Clorex disinfectant wipe that I waved inside the SAB before working. I think it helped with my trich problem as this last mini-SAB work I did, i used no chemicals and just sterile water on a paper towel and the trich came back. My next step is experimenting with spray Clorox.
The idea of the small glovebox is that (1) its easy to disinfect because of its small size and (2) it should stay clean since its sealed. So even thou the taped long-sleeves gloves create lots of turbulence inside the small glovebox, it wont matter if the spores have been killed with a can of disinfectant aerosol spray such as Clorox Aerosol spray.
Clorox Aerosol Spray
So the process will be open your small glovebox, insert all of your items that you need including a small can of disinfectant aerosol spray, then close the box up. Before doing any work inside the glovebox, spray the whole glovebox with the disinfectant aerosol spray. Wait for the aerosol spray to settle then do your mycology work. A good mold spore is a dead mold spore.
You know those Sci-Fi Movies where the person steps into the decontamination chamber? Same concept with this Mini Glovebox Decontamination Chamber or MGBDC 
Im pretty confident this method will reduce contaminations with converted mini-SABs.
-------------------- "What you must understand is that your physical dimension affects everyone in the higher dimensions as well. All things are interconnected. All things are One. Therefore, if one dimension is broken or out of balance, then all other dimensions will experience repercussions." - Pleiadian Prophecy 2020 The New Golden Age by James Carwin PROJECT BLUE BOOK ANALYSIS! (312 pages!) | Psychedelics & UFOs | Ready to Contact UFOs? | The Source on Mushrooms | Trippy Gematrix | Dj TeknoLogical | Fentanyl Test Kits R.I.P. Big Worm || The Start of the Ascension Process was 2020. Welcome to the Next Great Era of Earth 🌎🌍🌏
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elasticaltiger
Like Tigers in Coitus




Registered: 06/24/13
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Re: SAB contamination, how? M.G.B.D.C. - an experimental suggestion [Re: LogicaL Chaos]
#28648364 - 02/06/24 02:10 AM (3 months, 9 days ago) |
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Takeatrip and Logical, do either of you store your SABs on the floor when you're not using them?
-------------------- First time growing cakes? DON'T make a Shotgun Fruiting Chamber The Shmuvbox. - The Old TC's Like it Afraid to Start Growing From Your Own Prints? Drop it Like a Tiger! No Pouring. No Syringes. No Cutting. No flaming. No Contamination. No Bullshit. "The best thing to do while your waiting is to start more stuff. I usually got so much happening that I have tossed projects simply because I didn't have time for them. -Pastywhite QFT Pastywhite's Easy Agar Tek (PastyPlates) Tiger Drop Video Demos By munchauzen Van Gogh would’ve sold more than one painting if he’d put tigers in them.―Bill Watterson EZEKIEL 23:20
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LogicaL Chaos
Ascension Energy & Alien UFOs




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Re: SAB contamination, how? M.G.B.D.C. - an experimental suggestion [Re: elasticaltiger]
#28648367 - 02/06/24 02:13 AM (3 months, 9 days ago) |
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I keep mine at either 3ft off the floor or on a shelf about 5 ft off the floor.
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Skropi
Space Ritual


Registered: 11/01/23
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Re: SAB contamination, how? M.G.B.D.C. - an experimental suggestion [Re: LogicaL Chaos] 1
#28648578 - 02/06/24 07:42 AM (3 months, 9 days ago) |
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I don't know mate, I mean, my experience is very limited, but I don't think that trying to truly sterilise a SAB is a good idea. It just goes against the whole point of a SAB, based on everything I've read this far. When I poured my first 30 Petris dishes, I got more than half contaminated, and I had absolutely no idea why. The second time, I made 6 transfers, without using the "clam" technique, just open dishes, and got 100% success. The only thing I did differently was that I allowed more time for everything to settle in the SAB after I sprayed some soap water, I wore a mask, moved as slow as a platypus, and most importantly, I believe, I made sure that nothing would get above the dishes when they were open. Oh, and I used a cookie rack as a platform. It is very possible that I just got lucky though... I would definitely love for your idea to work out though, as it would mean a much less nerve wracking experience when working in the SAB. I mean, I need to calm myself after a session, it's a very intense experience, trying to avoid invisible enemies 🤣
-------------------- Ἄνδρα μοι ἔννεπε, Μοῦσα, πολύτροπον
Edited by Skropi (02/06/24 07:43 AM)
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Takeatrip
Stranger
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Re: SAB contamination, how? M.G.B.D.C. - an experimental suggestion [Re: elasticaltiger] 1
#28648964 - 02/06/24 12:39 PM (3 months, 9 days ago) |
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Quote:
elasticaltiger said: Takeatrip and Logical, do either of you store your SABs on the floor when you're not using them?
Mine is in my loft space, its boarded but not insulated. Its about 3 foot off of the floor now, however it was only about 6 inches or so before on some thick polystyrene sheets.
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