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Amazon Shop for: Agar, Petri Dish

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OfflineRohypnol
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Registered: 08/04/03
Posts: 267
Last seen: 1 year, 3 months
Using a ring to seperate mycellium from contam on agar
    #2085491 - 11/09/03 12:09 AM (13 years, 27 days ago)

After reading this website: http://www.botany.utoronto.ca/ResearchLabs/MallochLab/Malloch/Moulds/Contamination.html

I found out that by using a ring glued to the bottom of a petri dish, you can take advantage of the fungi's "tunneling" abilities to seperate mycellium from bacteria.

Basically:

You glue the ring to the bottom of the petri dish
Fill the dish with agar as normal, inside and outside of the ring - make sure not to get the ring wet or completely covered with agar.
Innoculate your contaminated sample inside the ring.

The mycellium should tunnel (under?) the ring, but the contamination will remain inside.


I am not completely sure how to glue the ring to the bottom, while allowing the mycellium to pass under it, so if someone could read that website and explain it to me I would be very thankful.


Quote:


When a mould is contaminated with yeasts or bacteria, dilution techniques usually work well. At times, though, yeast or bacterial cells so outnumber the spores of the desired fungus that dilutions are useless. Then it may be helpful to take advantage of the tunnelling or bridging abilities of fungus filaments. A technique sometimes used is to embed glass rings in the medium and surround the contaminated colony. A sterile glass ring, about 10 mm high and 15 mm in diameter, with three "feet" 2-3 mm in diameter glued or welded to the rim on the bottom, is placed on the bottom of a Petri dish. Sterile medium is then poured in to the usual depth, being careful not to overtop or wet the glass ring. After the medium has solidified, the mixed colony is inoculated into the glass ring. The mould filaments will be able to tunnel down under the ring and back up to the surface, while the bacteria or yeast, lacking the forward pressure of hyphae, will be confined to the ring.





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InvisibleJoshua
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Registered: 10/27/98
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Re: Using a ring to seperate mycellium from contam on agar [Re: Rohypnol]
    #2085669 - 11/09/03 02:23 AM (13 years, 27 days ago)

The ring has feet that help it to stand off of the bottom. These feet help create arches along the circumference of the ring on the bottom side. The mycelium can grow through these arches.

Joshua



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Invisiblemicro
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Re: Using a ring to seperate mycellium from contam on agar [Re: Joshua]
    #2088848 - 11/10/03 10:24 AM (13 years, 26 days ago)

Neat -- I never knew about this. Does this help isolate it from other fungi, though (besides yeasts, of course?)

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OfflineCultyVader
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Registered: 03/13/03
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Re: Using a ring to seperate mycellium from contam on agar [Re: Rohypnol]
    #2098109 - 11/12/03 07:04 PM (13 years, 23 days ago)

Can anybody get a picture of this happening? I'm haqving a lot of trouble picturing it in my mind. . .


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Invisibledeanofmean
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Registered: 12/07/02
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Re: Using a ring to seperate mycellium from contam on agar [Re: Rohypnol]
    #2099655 - 11/13/03 01:49 AM (13 years, 23 days ago)

that is a grate idea .
one could glue a rubber O ring to the lid of a petri dish, then fill the dish so the O ring just touches the agar .
it would be tedious, but there would be a barrier between the inside and outside of the ring .


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Invisibleplura16
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Registered: 09/25/03
Posts: 574
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Re: Using a ring to seperate mycellium from contam on agar [Re: deanofmean]
    #2101603 - 11/13/03 02:12 PM (13 years, 22 days ago)

i agree -- it is a great and interesting idea... heres some thoughts i had though..

if you glued the o ring to the lid of the dish, then filled up very high.. and replaced the lid - so the agar toughed the ring, there may be a lack of oxygen inside the ring. also how do get the sample in there?


maybe the same o ring on the bottom of the dish in the agar with some epoxy spots on one side... that would act as feet and create the gaps beneath the agar that it needs?




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OfflineRohypnol
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Registered: 08/04/03
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Re: Using a ring to seperate mycellium from contam on agar [Re: plura16]
    #2102463 - 11/13/03 06:24 PM (13 years, 22 days ago)

This is a different take on basically the same concept
http://www.nansnook.com/archives/tek/isotek.html


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Invisibledeanofmean
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Registered: 12/07/02
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Re: Using a ring to seperate mycellium from contam on agar [Re: Rohypnol]
    #2103012 - 11/13/03 08:57 PM (13 years, 22 days ago)

i think  i like that method even better .  :cool:
a couple drops of diluted h2o2 might even help there .

plura16, you are probably wright .
after further thinking on this; what if you made the ring of Tyvek .
then the ring could be made thick enough so it compresses slightly when the lid is taped down . no need for glue .
punch holes or cut slots near the bottom . this would allow gas exchange and keep any stray spores contained, at least until the lid is opened .
should be easy to make too .


Edited by deanofmean (11/13/03 09:49 PM)


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Offlinefreebee
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Registered: 11/25/03
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Re: Using a ring to seperate mycellium from contam on agar [Re: deanofmean]
    #2174235 - 12/13/03 06:47 AM (12 years, 11 months ago)

I don't think it's necessary to glue the ring to the bottom of the dish. The original article seems to be saying there are "feet" glued to the bottom of the ring, but the ring is simply setting on the bottom of the petri dish.
Another method is to pour a second layer of agar over top of a contaminated culture, then snag a piece of the mycelium when it grows up through the 2nd layer and transfer it to a new plate.

Freebee


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