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m_jacobs
Man Whore
Registered: 03/11/11
Posts: 121
Loc: CA
Last seen: 11 years, 27 days
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Great experiment EvilMush! Thank you for taking the time to share everything.
I'll be glad to bet a print that all plates come out clean.

Reasoning: 1. Evilmush is running a flowhood which means the room he's working in will be lower in airborne contams then the normal household.
2. The scalpel and needle were both flame sterilized for the 1st round of test. Then re-sterilized and wiped with alchol. That means the only vectors would be the ambient air or from the wipes themselves.
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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Re: Flame Sterilization Experiment [Re: m_jacobs]
#15187838 - 10/06/11 05:37 PM (12 years, 8 months ago) |
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Note for plates C,D,E,F the flame sterilizing & wiping (or just wiping) was done outside the flow of sterile air.
You bring up some valid points tho. Time will tell what happens.
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pierced074
Stranger
Registered: 07/28/11
Posts: 215
Last seen: 11 years, 6 months
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i believe all will be same,,,,this may be a dumb question...but...would the result be more pertinent if the open air sterilization was done in a high contaminant area that way you know you are actually sterilizing contams off of the needle/scalpel?
if your open air is extremely sterile anyway..you wouldnt be likely to contam the dishes....because your work area may not be same as ppl who live in high contam environment aka older houses etc......again...may have been dumb question...be easy on me
-------------------- disclaimer I do not produce, ingest, obtain, or distribute any illegal substances. Any photographs are computer generated creations or sniped from the internet and are for the purposes of entertainment and my fantasy of being a mycologist ,
Edited by pierced074 (10/06/11 06:02 PM)
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The Influence
Free Sheeks



Registered: 03/30/11
Posts: 6,067
Loc: Not Wisconsin
Last seen: 8 months, 29 days
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Re: Flame Sterilization Experiment [Re: pierced074]
#15187964 - 10/06/11 06:04 PM (12 years, 8 months ago) |
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Evilmushrooms, did you open an alcohol swab fresh, dip a cotton ball in iso and use right away or was the swab exposed to open air for a period of time?
I was thinking that if there are endospores that cannot be killed through steam sterilizing that there will definitely be some that cannot be killed with just iso?
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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Quote:
The Influence said: Evilmushrooms, did you open an alcohol swab fresh, dip a cotton ball in iso and use right away or was the swab exposed to open air for a period of time?
I was thinking that if there are endospores that cannot be killed through steam sterilizing that there will definitely be some that cannot be killed with just iso?
For the first plate I opened the swap in front of the flow hood.
For every swab after that I opened the swab in the open air as most people would do while inoculating PF jars without a glovebox.
This is by no means a proper experiment with controls and every variable dialed in and exactly the same. I just want to see how likely contams are to show up when wiping down pre sterilized instruments with ISO, or in the case of E & F if you skip flame sterilization all together and just wipe down with ISO.
Pierced: Every place will have a different spore load, but to give some background my lab has wall to wall carpet.
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The Influence
Free Sheeks



Registered: 03/30/11
Posts: 6,067
Loc: Not Wisconsin
Last seen: 8 months, 29 days
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Ya I know its not hard evidence, just wondering and trying to think of how variables will effect the outcome. Still a good experiment
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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To get hard evidence you would have to design an experiment, get 10-15 separate people to perform the procedures 100% accurately, collect data and compare.
A friend suggested an accurate way to get data would be to teach three different classes of high school kids how to grow edible mushrooms, get one class to flame, one class to flame and wipe, and the last class to only wipe. Collect data and compare contamination rates.
Seeing as I am a busy person and have a lot on the go I figured messing around with a few extra plates might give an idea of the likely hood of contaminates entering through the vector of cooling your instruments with an ISO swab.
I am not sure how things will turn out, all 6 plates very well may not contaminate at all.
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igigi



Registered: 09/10/11
Posts: 154
Loc: Sumer, Mesopotamia
Last seen: 4 years, 9 months
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Quote:
The Influence said: Evilmushrooms, did you open an alcohol swab fresh, dip a cotton ball in iso and use right away or was the swab exposed to open air for a period of time?
I was thinking that if there are endospores that cannot be killed through steam sterilizing that there will definitely be some that cannot be killed with just iso?
If I recall correctly, iso alcohol kills bacteria when drying off a surface. So a wet cotton ball could contain some bacteria, until it dried out?
-------------------- I --REFUSE-- TO BELIEVE Boeing 767s and WTCs are made out of exploded and unexploded nano-thermitic material. Google: bentham science thermitic. Click and READ the first link. I triple (WTCs 1, 2, and 7..) dog dare you. http://ae911truth.org/
Edited by igigi (10/06/11 06:57 PM)
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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Re: Flame Sterilization Experiment [Re: igigi]
#15188280 - 10/06/11 07:02 PM (12 years, 8 months ago) |
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When I say ISO swab, I am talking about the individually packaged piece of gauze soaked in 70% ISO, not a cotton ball.
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pierced074
Stranger
Registered: 07/28/11
Posts: 215
Last seen: 11 years, 6 months
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i applaud this experiment and in no way was i questioning you...and as you stated having wall to wall carpet that is very helpful info..as i have carpeted working area also.......excited to see the results cuz i have been doing open air needle into alcohol soaked sponge
-------------------- disclaimer I do not produce, ingest, obtain, or distribute any illegal substances. Any photographs are computer generated creations or sniped from the internet and are for the purposes of entertainment and my fantasy of being a mycologist ,
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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Re: Flame Sterilization Experiment [Re: pierced074]
#15188406 - 10/06/11 07:20 PM (12 years, 8 months ago) |
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I am in no way thinking that anyone is questioning me, I appreciate the input so that perhaps at a later time I can perform a more accurate experiment with more data points, or even someone else can take up the cause.
To get any level of accurate data I would think you would need to do at least 50 or so petri dish's for each method under the same strict procedures.
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TranscendingLife
I Don't Need a Life to Live



Registered: 06/09/10
Posts: 21,627
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you can sign me up for the next experiment once the flowhood is done. My house was built prior to 1930.
Also, just a tid bit of info, I have yet to have a contam from flaming the needle then wiping w/iso.
On the other side, I have had contams due to improper sterile procedure.
-------------------- AMU: We Quickly Answer Questions Here "One must accept the probability of failure to experience the elation of success." - TranscendingLife “A man of genius makes no mistakes; his errors are volitional and are the portals of discovery.” - James Joyce
      How I Do EVERYTHING      "Your vision will become clear only when you can look into your own heart…. Who looks outside, dreams; who looks inside, awakes."- Carl Jung "Anything that can be done chemically can be done by other means."- William S. Burroughs "You are as dead now as you will ever be" - Seth
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Cid2112
Newb



Registered: 09/24/11
Posts: 58
Loc: Ohio
Last seen: 5 years, 8 months
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Looking forward to the outcome on this
-------------------- Cid
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Samuel L Jackson
Bad Motherfucker


Registered: 12/10/09
Posts: 8,395
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there is no difference.
all you have to do to completely kill everything on a metal object is get it red hot. if wiping with alcohol was necessary you would see people in microbiology labs doing that shit.
they dont. you just heat your inoculation loop or whatever it is you want sterilized until red hot and then let it cool down.
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rkmate
Advocate

Registered: 10/07/11
Posts: 22
Last seen: 6 years, 4 months
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I think its all the same, isopropyl alocohol will do the same as a flame
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ky_distiller
Master Distiller




Registered: 06/16/11
Posts: 516
Loc: Kentucky
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Waiting with a worm on my tongue...
...baited breath.
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ely2121
Vagabond

Registered: 05/02/11
Posts: 385
Loc: 5th dimension
Last seen: 3 years, 5 months
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Re: Flame Sterilization Experiment [Re: ky_distiller]
#15194716 - 10/08/11 03:40 AM (12 years, 8 months ago) |
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 Thanks for doing this...for science!
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Wing
The Eye Tyrant



Registered: 04/25/11
Posts: 3,293
Last seen: 10 months, 6 days
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Re: Flame Sterilization Experiment [Re: ely2121]
#15195141 - 10/08/11 08:54 AM (12 years, 8 months ago) |
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If i had the resources i would do this without a flowhood. I bet a flowhood or even just a hepa filtered room could make all the difference.
-------------------- My Old Grow Logs
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FractalXplora
Grainiack




Registered: 02/11/06
Posts: 2,494
Loc: UK
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Re: Flame Sterilization Experiment [Re: Wing]
#15195235 - 10/08/11 09:22 AM (12 years, 8 months ago) |
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nice experiment EM666 looking forward to your results, cheers for taking the time to share
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EvilMushroom666
Heretic




Registered: 11/18/09
Posts: 10,289
Loc: Canada
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So far no signs of any contaminate bacterial or otherwise on any of the plates. It is still early though.
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