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Offlineimnorml
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Alcohol lamp outside or inside of still air box?
    #11319690 - 10/25/09 06:57 PM (11 years, 6 months ago)

I'd read in countless places by some of the most reliable sources to keep the sterilizing flame outside of the still air box and always found this confusing.

The motion of an arm exiting an arm hole MUST cause air outside the box to rush in since the volume of air plus volume of objects inside the box must remain constant, and removing your arm must mean that air equal in volume to your arm would have to rush in to replace it (some of this air will come in through the filter if there is one, but more will come in through the unimpeded arm hole).  This causes the problems of 1)dirtier air coming into the box and 2)stirring the air already in the box and thereby stirring up settled contams in the box.

On the other hand, having a lamp burning in the corner of the box would also cause air flow within the box due to the hot air rising.  However, I can't see the turbulence of the lamp being greater than the turbulence caused by an arm moving in or out.  It may be equivalent to the turbulence of arms and equipment moving around within the box, which happens regardless of where the lamp is.

I've read about the lamp using up the oxygen in the box, thereby drawing in air from the outside, but this doesn't make sense since the lamp replaces the O2 with CO2 and water vapor that it creates during combustion, so the chemical composition of the air would change but not the volume.  No change in volume = no total negative pressure = no air drawn in. (There is a negative partial pressure of oxygen, but it would be matched by the positive partial pressures of the other gasses, so no mass airflow would take place)

In either case, the box is keeping contams from dropping directly down from above and keeping the air more still than ambient air, which seems to be its most important functions, and is why it works pretty well regardless of where the lamp is placed.  I'm not arguing that having the lamp outside is not a sterile technique, I'm arguing that having the lamp inside improves an already sterile technique.

There are other benefits to having the lamp outside like being able to use alcohol/aerosols inside safely, no melting plastic, less space used, and not making the box uncomfortably warm, all of which would have to be weighed against any gains in sterility.

I know that this discussion has come up alot over the years, but I thought I could add to the conversation.  Thoughts?  Criticisms?


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OfflineMushface
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Re: Alcohol lamp outside or inside of still air box? [Re: imnorml]
    #11319780 - 10/25/09 07:12 PM (11 years, 6 months ago)

Could you just put a bic lighter in the glove box?


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OfflineThyrax
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Re: Alcohol lamp outside or inside of still air box? [Re: Mushface] * 1
    #11319808 - 10/25/09 07:16 PM (11 years, 6 months ago)

glove box + alcool lamp inside = boom

or at least its not a good idea, just keep it outside.


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Offlinemushking
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Re: Alcohol lamp outside or inside of still air box? [Re: Thyrax]
    #11319896 - 10/25/09 07:30 PM (11 years, 6 months ago)

its not that hard to not get contaminations, your over thinking, dont worry about it.

fuck the contamination nazi's.


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OfflineBrennus
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Re: Alcohol lamp outside or inside of still air box? [Re: mushking]
    #11319967 - 10/25/09 07:43 PM (11 years, 6 months ago)

Always keep your alcohol lamp outside your glove box.

Tight space + alcohol fumes + fire =  :onfire:


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OfflineSN122A
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Re: Alcohol lamp outside or inside of still air box? [Re: Brennus]
    #11320095 - 10/25/09 08:02 PM (11 years, 6 months ago)

I guess I have always used the glove box with the frame of mind that its intended function is to minimise the movement of air.  When you push/pull your arms the displacement will create a sudden and temporary movement of air then it will settle as opposed to a flame creating continuous circulation of the air while its burning…

For instance I will keep a bottle of alcohol/bleach inside the glove box and after I place my arms in spray liberally over the petri’s, jars (with foil still on) etc so the moisture of the mist will bind to the particles both sterilising and pulling them onto the floor of the glove box.

Many people (I think Stamets mentioned it as well) suggest that this way of removing airborne contaminants via moisture is very efficient and liken it to the clean/fresh air after if has rained extensively…

Works for me!  Anyone else do this?


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OfflineMushface
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Re: Alcohol lamp outside or inside of still air box? [Re: SN122A]
    #11320138 - 10/25/09 08:12 PM (11 years, 6 months ago)

For me personally I'm following rr's videos to a T.


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OfflineDamion5050
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Re: Alcohol lamp outside or inside of still air box? [Re: Brennus]
    #11320148 - 10/25/09 08:14 PM (11 years, 6 months ago)

Quote:

Brennus said:
Always keep your alcohol lamp outside your glove box.

Tight space + alcohol fumes + fire =  :onfire:





This is very true, so don't do it.  About 4 days ago some one did this exact thing and blew up his glove box.


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Offlineimnorml
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Re: Alcohol lamp outside or inside of still air box? [Re: imnorml]
    #11320205 - 10/25/09 08:24 PM (11 years, 6 months ago)

Any of the fumes from the lamp get burned as they pass through the hole with the wick (my lamp is sealed everywhere else).  They can't get into the rest of the box without passing through the fire, which they can't do... because they're flammable.  If you sterilize with alcohol, I imagine it could be a problem.  Explosions happen when you allow an area to get filled with fumes then introduce fire or a spark.  I do love that guy-on-fire animation, though.  I like the bleach/alc spray idea.  Good results with it SN?


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Offlineimnorml
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Re: Alcohol lamp outside or inside of still air box? [Re: imnorml]
    #11320284 - 10/25/09 08:35 PM (11 years, 6 months ago)

Damion, was the person using alcohol wipes or just a lamp?


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OfflineDamion5050
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Re: Alcohol lamp outside or inside of still air box? [Re: imnorml]
    #11320331 - 10/25/09 08:43 PM (11 years, 6 months ago)

Quote:

imnorml said:
Damion, was the person using alcohol wipes or just a lamp?





Not sure I'll try and find the post.


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OfflineDamion5050
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Re: Alcohol lamp outside or inside of still air box? [Re: Damion5050]
    #11320339 - 10/25/09 08:44 PM (11 years, 6 months ago)



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Offlineimnorml
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Re: Alcohol lamp outside or inside of still air box? [Re: Damion5050]
    #11320401 - 10/25/09 08:58 PM (11 years, 6 months ago)

Quote:

Damion5050 said:
http://www.shroomery.org/forums/showflat.php/Number/11310629




seems that this person had an open container of alcohol in his GB and introduced flame.  Obviously very bad idea.  Thanks for finding the post.

The first post in this thread was inspired by me using an alcohol lamp outside of the still air box a la RR's instruction.  It was while following the instructions that the problems with the procedure occurred to me. Only did it today, so I don't know how it turned out yet... I'll get back to you in a few days. 

Haven't yet tried the lamp in the GB, but perhaps I'll try a BBQ lighter instead, one with a trigger away from the flame so I don't end up with burnt latex stuck to me.  This will solve most of the problems with the alcohol lamp (air disturbance will only last for a few seconds instead of continuously, no explosion risk as long as I use 10% bleach solution instead of aerosols and alcohol to disinfect).


Edited by imnorml (10/25/09 08:59 PM)


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OfflineBreakfast Crew
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Re: Alcohol lamp outside or inside of still air box? [Re: imnorml]
    #11320501 - 10/25/09 09:11 PM (11 years, 6 months ago)

You have got to be kidding me, no one is that retarded.  You have some book smarts (Sorta) but absolutely no common sense.

Still air boxes are silly, you want an airtight container.  A little bit of silicone bought from any hardware store goes a long way.

But I lol at people freaking out over sterility issues. :2cents:


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Offlinebeto
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Re: Alcohol lamp outside or inside of still air box? [Re: Breakfast Crew]
    #11320801 - 10/25/09 10:12 PM (11 years, 6 months ago)

Always outside!

You don't need to "over worry" that much, as stated above. If u follow the basic sterile procedures youre fine. Wash your hands and forearms, use a mask and latex gloves, alcohol and that's pretty much it.

This is working for me and i'm new at this so, it's not that hard. I use a still air box for everything,  i poured over 60 petri dishes already  only 3 or 4 got contaminated so far.


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OfflineRogerRabbitM
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Re: Alcohol lamp outside or inside of still air box? [Re: imnorml]
    #11321610 - 10/26/09 12:22 AM (11 years, 6 months ago)

Quote:

imnorml said:

The motion of an arm exiting an arm hole MUST cause air outside the box to rush in since the volume of air plus volume of objects inside the box must remain constant, and removing your arm must mean that air equal in volume to your arm would have to rush in to replace it




Correct, but you're not actively inoculating or opening a petri dish while you're removing your arm to flame sterilize.  Once you re-insert your arm, the air slows down.  Remember, we don't have sterile air in a box, it's still air.


Quote:

imnorml said:
On the other hand, having a lamp burning in the corner of the box would also cause air flow within the box due to the hot air rising.  However, I can't see the turbulence of the lamp being greater than the turbulence caused by an arm moving in or out. 



The flame causes far more turbulence than your hands do, and the O2 being consumed by the flame must be replaced by air from outside moving into the box, thus causing disturbed, turbulent air, not still air. 

In addition, the heat produced by the flame will cause condensation on the walls of the box, making it hard to see.  The heat will also rapidly turn the box into an oven.  Even with my alcohol lamp outside the glovebox, it's not unusual to see a 10F rise in temperature in the room I'm doing sterile work, simply from the heat of the flame.

I strongly recommend keeping the flame on the outside.
RR


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Offlineimnorml
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Re: Alcohol lamp outside or inside of still air box? [Re: RogerRabbit] * 1
    #11323315 - 10/26/09 12:03 PM (11 years, 6 months ago)

You guys have definitely talked me out of using an alcohol lamp in the box.  Thanks for the input.  I'm still convinced that using a lighter inside the box would be preferable to using an alc lamp outside, though.

Quote:

RogerRabbit said:
Correct, but you're not actively inoculating or opening a petri dish while you're removing your arm to flame sterilize.  Once you re-insert your arm, the air slows down.  Remember, we don't have sterile air in a box, it's still air.





I really don't see why the air would calm down upon reinsertion of your arm, it would likely be equal to the disturbance caused by removing it and is analogous to the initial insertion of jars/dishes/equipment which most sources say requires a 5 minute period of waiting while the air calms down.

The air would calm down pretty fast in such a confined place anyway, which is why the box is effective regardless of how you use it, but I'm convinced that it is more effective without the arm moving in and out.

Quote:

RogerRabbit said:
The flame causes far more turbulence than your hands do, and the O2 being consumed by the flame must be replaced by air from outside moving into the box, thus causing disturbed, turbulent air, not still air. 





I think that you're right about the constant turbulence caused by an alcohol lamp due to rising air, which makes it a bad idea.  A lighter, though would still work since you're "not actively inoculating or opening a petri dish" while you're flame sterilizing.

A lighter also wouldn't run long enough to cause a great temp increase or condensation.

The idea of "air" needing to replace the O2 burned is not true (although it doesn't matter since we're not worried about sterile air).  Consuming O2 in the box means that more O2 will flow into the box due to the partial pressure effect.  In statistical mechanics this means that since there is a higher concentration of O2 outside than inside it is more likely for an O2 molecule to move in than out.  However, the opposite is true for the CO2 and H2O vapor, thereby balancing the pressure.  As long as the total number of air molecules in the box doesn't change there is no net negative pressure.

Quote:

Wikipedia said:
In a mixture of ideal gases (or real gasses at normal ambient conditions), each gas has a partial pressure which is the pressure which the gas would have if it alone occupied the volume. The total pressure of a gas mixture is the sum of the partial pressures of each individual gas in the mixture.





A mold spore (which is 1000-100,000 times the size of an O2 molecule) sitting at the the threshold of the box is clobbered inward by the more plentiful O2 molecules on the outside while simultaneously being clobbered outward by the more plentiful CO2 and H2O inside.

Next time I use my box, I'll sterilize everything with 10% bleach and use a lighter inside.  If I see an increase in contams I'll post about it.


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OfflineRogerRabbitM
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Re: Alcohol lamp outside or inside of still air box? [Re: imnorml]
    #11323352 - 10/26/09 12:09 PM (11 years, 6 months ago)

Quote:

The idea of "air" needing to replace the O2 burned is not true (although it doesn't matter since we're not worried about sterile air).  Consuming O2 in the box means that more O2 will flow into the box due to the partial pressure effect.  In statistical mechanics this means that since there is a higher concentration of O2 outside than inside it is more likely for an O2 molecule to move in than out.  However, the opposite is true for the CO2 and H2O vapor, thereby balancing the pressure.  As long as the total number of air molecules in the box doesn't change there is no net negative pressure.




Irrelevant.

The O2 being consumed by the flame must be replaced.  Thus there is turbulence and air movement in and out of the box, which is what you don't want. The heat also causes convective turbulence, again stirring the air. Nobody is counting molecules.
RR


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Offlineimnorml
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Re: Alcohol lamp outside or inside of still air box? [Re: RogerRabbit]
    #11323457 - 10/26/09 12:33 PM (11 years, 6 months ago)

Gas molecules always move around constantly, in every direction, and very quickly.  What we are worried about is mass movements of molecules.  Partial pressures don't cause mass movements of molecules as long as there is no total pressure differential.

Total number of particles is directly related to pressure.  When you measure pressure you are counting molecules in a sense.

I do agree that the heat from the flame will cause turbulence, still less than the moving arm, though, I'll guess, as long as the flame is short lived.  I'll test it with some incense smoke inside the box so I can see the air movement.  I'll get back to you on how it turns out.


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Offlineashash1212
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Re: Alcohol lamp outside or inside of still air box? [Re: imnorml]
    #11582353 - 12/04/09 09:05 AM (11 years, 5 months ago)

Theres a general consensus that anything spoken by RR is considered mushroom law. disobeying the law can result in contamination and then everyone loses which is what we dont want, so take his advice.


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