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Offlinesagecl

Registered: 06/18/18
Posts: 18
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Working under alcohol lamp
    #25334684 - 07/18/18 11:02 PM (5 years, 6 months ago)

Any thoughts on this technique? I know it's not as good as a proper SAB but it is very oftenly used on real microbiology labs.
Is it fittable to agar duty ?


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InvisiblebodhisattaMDiscordReddit
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Re: Working under alcohol lamp [Re: sagecl] * 1
    #25335135 - 07/19/18 09:08 AM (5 years, 6 months ago)

Its supposed to be a bunsen burner not an alcohol lamp. And i wouldn't suggest it to more modern technique. It's ok for basic bacteriological work for highschool and college kids or basic QA work for a small lab. We use petri dishes as inoculant, rather than just diagnostic tools. Thus we require completely aseptic plates.


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OfflineSolipsis
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Re: Working under alcohol lamp [Re: bodhisatta]
    #25335344 - 07/19/18 11:44 AM (5 years, 6 months ago)

IIRC one advantage of a burner is that it creates an air current which goes upwards plus pulls in air which gets sterilized from the intense heat.

I don't have a flowhood but I have tried using an alcohol burner in a glovebox once or twice which is a terrible idea. What I do now is I have a glovebox with a really small opening / slot in one side and the glovebox sits on my kitchen counter not far away from the stove.

So I use the stove to sterilize tools which I pass through the slot into the glovebox onto a piece of natural stone for it to cool. The bonus is that it draws away air which gets sterilized and the airflow is less likely to be compromising.

The glovebox is large enough to be comfortable so I really don't miss that it's not a SAB, esp with things having a standard place in the box.

Sorry I misunderstood the OP a bit... I recall a bit of that in biology back in college but I really don't recommend it as a substitute for a box or hood. I don't think a box is too expensive to make.


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OfflineFishLevelMidnight
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Re: Working under alcohol lamp [Re: Solipsis] * 1
    #25355985 - 07/30/18 01:36 PM (5 years, 5 months ago)

In my new set up I have a propane tank and 2 big Bunsen burners. I've poured plates, inoculated sterile bags with grain, did g2g transfers, inoculated LC, make syringes...
Everything I had been doing at work with the flowhood I am doing with the sterile field created by the flame.

Took a little bit of tinkering to figure out how to maneuver plates and things to be workable and in the sterile field but I will probably put off building an at home flow hood for a little while as this is working great for me. Spent many hours witht he flame running and my tank is still pretty heavy.

I think Josex also recently moved to this set up with success, so as long as you know what you're doing, know how to pick out contaminants and are willing to have a little % of your projects contam (I' at maybe 5% or so) it is a viable method.

I would suggest against an alcohol lamp though.


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InvisibleJosex
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Re: Working under alcohol lamp [Re: sagecl]
    #25358440 - 07/31/18 02:39 PM (5 years, 5 months ago)

Quote:

sagecl said:
Any thoughts on this technique? I know it's not as good as a proper SAB but it is very oftenly used on real microbiology labs.
Is it fittable to agar duty ?




Get yourself a pair of bunsens, alcohol lamp aint gonna cut it. My success rate with the bunsens is 100% and besides G2G I've done everything without a hitch, you just need to know how they work and it's actually really hard to get contams once you know what you're doing.

The reason people haven't tried this before for myco escapes me, but it certainly isn't less aseptic than a ghetto box and still air.

I actually started experimenting with fire because the place I grow in is mold city and I had issues working with the SAB. I'd disregard the fuck out of any opinion from people who haven't tried this. :shrug:


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Invisible36fuckin5
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Re: Working under alcohol lamp [Re: Josex]
    #25369677 - 08/06/18 08:51 AM (5 years, 5 months ago)

I just don't see the advantage of working under a burner. You're gonna have a tiny workspace which is gonna up the time it takes to get shit done. You're also gonna have to buy a Bunsen burner if you don't have one, plus you'll have to keep buying fuel. I'd rather spend $10-20 on a box and be able to fit a bunch of shit in it.


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Pat The Bunny said:
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bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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InvisibleJosex
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Re: Working under alcohol lamp [Re: 36fuckin5]
    #25370917 - 08/06/18 07:18 PM (5 years, 5 months ago)

You're only capable of doing one thing at a time in the SAB and you use a very small space for that, dont you.
Same thing with the bunsens, with 2 of them working at the same time you have a sizeable sphere where you can do your sterile work comfortably.

Jars are lined up on the table waiting their turn to be inoculated, it's by no means slower plus you don't have to set up a SAB nor do you need to move as carefuly as in the SAB, because the bunsens really allow more room for error or clumsiness, not to mention working open air trumps working in a cramped-up, uncomfortable SAB with poor visibility.

They also consume really little gas. I've been doing this for months and the gas cylinders are heavy as if I just bought them.


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OfflineMtez44
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Re: Working under alcohol lamp [Re: Josex]
    #25371774 - 08/07/18 06:53 AM (5 years, 5 months ago)

I have to say I am intrigued by this technique as opposed to a SAB. I fucking hate still air boxes. So uncomfortable and no matter how clear the plastic is I dont like the reduced visibility. Then you are supposed to move so damn slow. Just my opinion.

Fisherman/Josex can you recommend a burner? I was looking at the larger merker style ones on amazon but they were around $60 each


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OfflineFishLevelMidnight
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Re: Working under alcohol lamp [Re: Mtez44]
    #25371826 - 08/07/18 07:30 AM (5 years, 5 months ago)

I think I have the same style as Josex but I got for stupid cheap at the university (they have this awesome store that sells old equipment, science stuff as well as items from dorms and other parts of the campus), so I can't send you in the proper direction of a source :frown:


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InvisibleJosex
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Re: Working under alcohol lamp [Re: FishLevelMidnight] * 1
    #25371950 - 08/07/18 08:41 AM (5 years, 5 months ago)

Ya Mekers are at least twice as expensive as bunsens, but they're so badass and shiny. I got my 2 bunsens from Ebay and other spanish site, 30 euros each.


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InvisibleChemBioWiz
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Re: Working under alcohol lamp [Re: sagecl]
    #25411262 - 08/24/18 10:43 PM (5 years, 5 months ago)

Thanks for posting this question man!

I use a Bunsen burner frequently for pouring plates, making transfers, cloning, inoculating jars, and doing other sterile work. This technique seems to get a lot of hate around here, but it's reliable, cheap, and very effective if performed correctly. Honestly, I'm surprised more new folks don't at least try it out since it's so accessible, especially considering that there's just as much room to maneuver as there is in your average glove-box/SAB. It's definitely much cheaper than building a flow hood, although it does have its obvious drawbacks.

Just like most techniques, when people mess this one up, it usually comes down to sloppy procedure -- they don't have a strong enough updraft (e.g. their flame isn't "jetting"), they're working in an area with a lot of competing air currents, they're breathing heavily downward into their workspace, or they work too far outside the updraft cone diameter (it's only about 60 cm (24 in) maximum).

I'll take a few pictures or make a video either this or next weekend and post the technique I use if you folks are interested. I don't particularly feel like starting an argument with people over this since a lot of you seem to have a strong opinion about it, but I think it would be good to at least showcase it as a capability in our toolbox.


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InvisibleJosex
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Re: Working under alcohol lamp [Re: ChemBioWiz]
    #25412143 - 08/25/18 09:50 AM (5 years, 5 months ago)

Well put. :mostinteresting:


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OfflineDutchMyco
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Re: Working under alcohol lamp [Re: Josex]
    #25412380 - 08/25/18 12:00 PM (5 years, 5 months ago)

Used it for agar work many years ago, not for mushroom cultivation though so can't really comment on that.
Some weeks ago I came across a technique used for sterile work for growing orchids, which used working on a screen above a boiling pot of water. Not sure if this is in any way more practical than a SAB, but I found it creative enough to share here.


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Offlinecilasiah
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Re: Working under alcohol lamp [Re: ChemBioWiz]
    #25412456 - 08/25/18 12:38 PM (5 years, 5 months ago)

Yes please! Make a tutorial for your techique! will be waiting!
thanks


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Invisible36fuckin5
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Re: Working under alcohol lamp [Re: cilasiah]
    #25412683 - 08/25/18 02:37 PM (5 years, 5 months ago)

Working over a boiling pot of water is dangerous and stupid. Steam can and will burn the shit out of you, not to mention it could melt your plates and cause all kinds of condensation.


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Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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InvisiblebodhisattaMDiscordReddit
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Re: Working under alcohol lamp [Re: 36fuckin5]
    #25414613 - 08/26/18 01:04 PM (5 years, 5 months ago)

Plant tissue culture doesn't demand nearly the same level of cleanliness as fungal tissue culture work.


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InvisibleWolfGang G
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Registered: 07/20/17
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Re: Working under alcohol lamp [Re: bodhisatta]
    #25415451 - 08/26/18 08:07 PM (5 years, 5 months ago)

This is my take on using the burner. It will work well with a laminar flow hood, but in a still air environment it causes some heat convection and takes time to complete the process. It can be done will a still air box with great success though.

This is how my evolution of culture has taken me. I use disposable scalpels (#11), they cost about $5 for 10 of them and they make it through the pressure cooker. I keep 30 rolled up in foil and undo the foil as I need another scalpel. This technique takes away a lot of risky movements and takes less time to complete. You never have to leave the still air box because everything is already sterile.


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Invisible36fuckin5
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Re: Working under alcohol lamp [Re: WolfGang G]
    #25415670 - 08/26/18 09:42 PM (5 years, 5 months ago)

Quote:

WolfGang G said:
This is my take on using the burner. It will work well with a laminar flow hood, but in a still air environment it causes some heat convection and takes time to complete the process. It can be done will a still air box with great success though.

This is how my evolution of culture has taken me. I use disposable scalpels (#11), they cost about $5 for 10 of them and they make it through the pressure cooker. I keep 30 rolled up in foil and undo the foil as I need another scalpel. This technique takes away a lot of risky movements and takes less time to complete. You never have to leave the still air box because everything is already sterile.




Did you even read the thread?


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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Offlinesagecl

Registered: 06/18/18
Posts: 18
Last seen: 1 year, 4 months
Re: Working under alcohol lamp [Re: Josex]
    #25418232 - 08/27/18 11:47 PM (5 years, 4 months ago)

Well thanks everyone for the answers, got myself a bunsen very cheaply from aliexpress, now i'm evaluating ways to connect it to gas tank.
I know a bunsen won't replace a sab or a fh, but for small work it could be handy and I think is good lab practice.

Quote:

Josex said:
Get yourself a pair of bunsens, alcohol lamp aint gonna cut it. My success rate with the bunsens is 100% and besides G2G I've done everything without a hitch, you just need to know how they work and it's actually really hard to get contams once you know what you're doing.

The reason people haven't tried this before for myco escapes me, but it certainly isn't less aseptic than a ghetto box and still air.

I actually started experimenting with fire because the place I grow in is mold city and I had issues working with the SAB. I'd disregard the fuck out of any opinion from people who haven't tried this. :shrug:



Thanks for the reply man, I've followed your teks and just love them, and the fact that you're not from the us really helps since i'm not either, so you kinda understand the low availability of some items outside us.


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Offlinesagecl

Registered: 06/18/18
Posts: 18
Last seen: 1 year, 4 months
Re: Working under alcohol lamp [Re: WolfGang G]
    #25418237 - 08/27/18 11:53 PM (5 years, 4 months ago)

Quote:

WolfGang G said:
This is my take on using the burner. It will work well with a laminar flow hood, but in a still air environment it causes some heat convection and takes time to complete the process. It can be done will a still air box with great success though.

This is how my evolution of culture has taken me. I use disposable scalpels (#11), they cost about $5 for 10 of them and they make it through the pressure cooker. I keep 30 rolled up in foil and undo the foil as I need another scalpel. This technique takes away a lot of risky movements and takes less time to complete. You never have to leave the still air box because everything is already sterile.




You could consider buying a metallic scalpel and the blades separately, the grip of the scalpel cost me like 2$ and each blade around  .2$ each, so I just throw them away after every time. I think nothing in that sab is actually sterile, it's just that contams get stuck onto the walls of everything and they are not floating around, so they don't enter your work.


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