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OfflinePTreeDish
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Registered: 04/22/18
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Carbon Build Up on Flame Sterilized Scalpel
    #26492646 - 02/18/20 11:12 PM (3 years, 11 months ago)

I use these #10 sterile scalpels for transfers and one of these benzomatic propane torches for flame sterilization.

I've noticed that sometimes a black flakey material, maybe carbon, will build up on the blade after it has been heated red hot and flake off in the dish when cooled. I've noticed more contamination occurs when this happens but the sample size is small so take it with a grain of salt.

Has anyone had this happen or know how to prevent it? My hunch is that either I'm holding the scalpel in the wrong part of the flame and/or the fuel isn't "clean" so it's building up as soot. Will be trying a denatured OH flame tomorrow.


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OfflinePTreeDish
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Re: Carbon Build Up on Flame Sterilized Scalpel [Re: PTreeDish]
    #26493691 - 02/19/20 04:33 PM (3 years, 11 months ago)

Any thoughts or feedback here?


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InvisibleSirSpanksAlot
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Re: Carbon Build Up on Flame Sterilized Scalpel [Re: PTreeDish]
    #26493985 - 02/19/20 07:37 PM (3 years, 11 months ago)

what flavor metal is it? if its steel like most common blades, its likely slag or iron oxides forming with a combination of your fuel. <metal impurities

the nitrogen in the air and the fuel are contaminating the steel most likely as you say you : youre likely holding your blade a bit too close to the fuel source? or too far.

theres a sweet spot when it comes to the cones of heat that a flame torch gives off (you want that bluish/white flame -inside that oxidizing flame, there is a sharp cone that you will try to keep your blade from touching but not outside of your blue cone) -torch welding videos can give you an idea if youre running a red/yellow fuel flame.

not assuming things but I hope this helps in a way. a tip that I use for keeping down the possibility of contaminates while also not cooking any clone slices that I take, is to have a small dish/vessel of H2O2 to quench my red hot blade, before taking cuts (all in the SAB; torch excluded)


Edited by SirSpanksAlot (02/21/20 01:19 AM)


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OfflinePTreeDish
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Re: Carbon Build Up on Flame Sterilized Scalpel [Re: SirSpanksAlot]
    #26494212 - 02/19/20 11:02 PM (3 years, 11 months ago)

That's really helpful. I linked to the blades I'm using - it doesn't say exactly the metal used, just "surgical blades". It is probably steel though.

Would switching to a denatured alcohol flame be of any help? I'm guessing it would burn cleaner but I'd still have to place the blade in the right position?

Also, re: quenching the blade in H202. Are you using sterile water? I'd have presumed that would introduce another vector for contams.


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OfflineMr.Wizard
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Re: Carbon Build Up on Flame Sterilized Scalpel [Re: PTreeDish]
    #26494364 - 02/20/20 03:20 AM (3 years, 11 months ago)

If I see any while I'm flaming I'll wipe on the alcohol paper towel, and reheat. I do an agar 'quench', just swiping it in the sides a few times. I haven't seen contams yet, but lifetime dish count is pretty low.

On a side note - I've made a forge (a few), and dabbled in blacksmithing. I always look forward to heating up my scalpel. I love that the verb 'quench' can be applied to mycology as well :rofl:


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InvisibleSirSpanksAlot
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Re: Carbon Build Up on Flame Sterilized Scalpel [Re: PTreeDish]
    #26496344 - 02/21/20 01:26 AM (3 years, 11 months ago)

H2O2 (hydrogen peroxide) I normally use straight out the bottle, wiped down in the SAB. beyond that, I may be introducing contaminates, but Ivea low percentile of failed cultures (im just a shite fruit cultivator).

Beyond the possible impurities on the blade,, I figure the corrosive chemical reaction of the H.peroxide is capable of detering most hitchhiking contam.s surviving the flame. Beyond that, there is just the chemical contamination, but we only worry for live contams rather than minute chems of spent fuel and nitrogen build up and oxides - the quench process tends to flake the oxides material off in the H2O2 or the agar

If it bothers you much, it is posibly cheaper and more easy to acohol burner flame than torching (i prefer torching since its convenient to see the flame and cherry red)


Edited by SirSpanksAlot (02/21/20 01:27 AM)


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OfflineMrBovineJoni
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Re: Carbon Build Up on Flame Sterilized Scalpel [Re: SirSpanksAlot]
    #26496387 - 02/21/20 02:48 AM (3 years, 11 months ago)

Quote:

SirSpanksAlot said:
detering most hitchhiking contam.s surviving the flame.

.......
(i prefer torching since its convenient to see the flame and cherry red)



That is the only good way to flame sterilise at home. If the scalpel blade glows red  - there cannot be any live/live contaminants left on the surface of the blade; no biological organism known to humans so far can survive that temperature (e.g. steel glow red temperature is around 900F/480C). So peroxide in this case adds absolutely nothing.

Wiping down the scalpel handle before flaming (the parts that are not flamed)is still beneficial but 70% isopropyl is much better for it than peroxide solution.


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OfflinePTreeDish
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Re: Carbon Build Up on Flame Sterilized Scalpel [Re: Mr.Wizard]
    #26497409 - 02/21/20 05:51 PM (3 years, 11 months ago)

Quote:

Mr.Wizard said:
If I see any while I'm flaming I'll wipe on the alcohol paper towel, and reheat. I do an agar 'quench', just swiping it in the sides a few times. I haven't seen contams yet, but lifetime dish count is pretty low.

On a side note - I've made a forge (a few), and dabbled in blacksmithing. I always look forward to heating up my scalpel. I love that the verb 'quench' can be applied to mycology as well :rofl:




That sounds cool. I'd totally buy a hand-forged mycology scalpel from you.

I did a few new transfers and cultures today and I went ahead and quenched in the agar of the clean plate. It made a quenching noise as the agar quickly steamed where I touched it.

Do you turn off your torch between each flaming or leave it on for the duration of your work? In spite of wiping down the torch vigorously, it is off to the side and I'm concerned touching it right after flaming but before culturing could introduce a new contam vector.


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InvisibleSirSpanksAlot
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Re: Carbon Build Up on Flame Sterilized Scalpel [Re: MrBovineJoni]
    #26497510 - 02/21/20 06:57 PM (3 years, 11 months ago)

I usually give a isopropyl/hydrogen peroxide mix, then shake up for the inside of the syringe between knocking up jars. Healthy squeezes with little to no contaminate problems so far (but I would appreciate a feedback on this -im nearly certain that I can cut onne out entirely) -i was under the impression isopropyl doesnt kill contams?



-----@  PTreeDish
if youre really good at keeping an open flame on at low settings, moving between work and the SAB shouldnt be much of a problem if thats your concern. Be wary that sudden uncalculated moves can be hazardous

-always keep your flame visible and face away so your needle points away, or you'll be shooting out liquids/vaporized mycelium (my experience is that I recycle syringes cause broke and cheap -plus the ole swish above was the method of sitting/soak for the majority of set up for working)


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