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Offlineadadada
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How to prevent condensation in agar plates
    #23640825 - 09/13/16 07:55 AM (7 years, 4 months ago)

Hey all,

I was frustrated with all of the water condensation I've been getting with my agar plates.

Since I use a pour method, I've been pouring the agar while still hot into the sterile plates and as they cool they give off a lot of vapor which condenses on the lid of the plate.

While this is mostly an annoyance, the mobile water droplets can act as a vector for contamination to travel within the plate.  The moisture can also form an airtight seal of liquid water around the edge of the plate, preventing gas exchange.  Not good.

So I started googling around and found a few tips which I wanted to share with you all ...

Prevention

These steps help prevent condensation if you haven't poured your plates yet.  Please note that these tips may not be applicable for no-pour methods.

  • Let your agar cool to 127-131F (53-55C) before pouring.  This will minimize evaporation but means you'll have to act fast once the agar reaches this temp since it will start to solidify at 122F (50C).  I like to fill the pressure cooker with water to the height of the agar and can then use an instant read thermometer to check the water temp around the agar jar before removing it ... bearing in mind that the agar will be slightly hotter than the water it's sitting in.  A hot water bath or well calibrated hot plate would be very useful to slow the cooling rate of the agar container and keep it in that critical 53-55 C range.
  • Stack the plates and put an empty plate on the top for additional insulation.  Stacking plates in this way slows the cooling process and encourages condensation on the sides rather than top or bottom of the plate.  This has sometimes helped me but seems less reliable. 
  • Leave the plates partially open until they solidify.  Not for the faint of heart! If you're using this method, I would pour extra plates to cover contamination losses, let the plates cool with some sterile cover above them (if using a still air box), and gestate the plates (after cooled and covered) for a few days  to see if any contamination is present before inoculating.  This should be very effective in reducing condensation but is time consuming and more prone to contamination.

Mitigation

If your plates are already poured and have condensation in them then you can do the following to mitigate the impact of having free flowing water in your plate ...

  • Store the plate upside down.  This prevents the water from contacting the agar and mycelium.  So if there is contamination present it should not be able to use the water movement to spread.
  • Temporarily remove the condensation by placing a warm object on the plate for 10-15 min.  I like to use a mug of hot coffee.  The gentle warmth from the cup will cause the water droplets to evaporate without harming the mycelium.  The condensation will unfortunately return but it will provide several minutes of clear viewing, which should be enough time to assess the plate.  Please note that this will not work if you're storing your plates upside down.

adadada


Edited by adadada (09/13/16 08:10 AM)


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Offlineadadada
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Re: How to prevent condensation in agar plates [Re: adadada]
    #23640865 - 09/13/16 08:18 AM (7 years, 4 months ago)

Here are a few other thoughts I had but have not yet tried.

Mitigation

  • Place the plates in a stream of air.  A small fan should do the trick.  The air movement should reduce the air pressure and increase evaporation ... possibly pulling the water vapor out of the plate.  This might increase contamination rates though.
  • Place the plates in an enclosed container with a desicant (e.g. dry white rice, those packets from electronics boxes, etc).  The desicant would reduce the relative humidity within the container and essentially pull the excess water out of the plate.  Would have to be careful not to dry out the plate(s) too much though.  Also, would need to make sure the desicant is active by heating it in a low oven for several hours before exposing it to the plates.


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Offlinekatbusa
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Re: How to prevent condensation in agar plates [Re: adadada]
    #23655346 - 09/18/16 11:05 AM (7 years, 4 months ago)

Condensation happens. You can help mitigate some of it by stacking the plates while the agar is hardening. In all condensation shouldnt hurt the plate. Another thing I do is once the agar is fully hardened ill tap the plate and force the condensation to make droplets. I then carefully dump out the excess water. This is all done under the flow hoot of course.

I have also noticed that over time the condensation goes away. However the parafilm I use to seal the plates is somewhat breathable. So that could be why condensation seems to go away for me.


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Offline50mmprime
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Re: How to prevent condensation in agar plates [Re: katbusa]
    #24553325 - 08/14/17 02:14 PM (6 years, 5 months ago)

I'd second that the two most important aspects are: 
1 - waiting for the agar to cool a bit before pouring
2 - stacking the plates while they cool

One more piece I've found helpful is shuffling the stack. While the plates are cooling, every few minutes switch the ones on the top for the ones on the bottom. I guess the idea is that the slower the plates cool, the more time there is for water vapor to diffuse out.

If you leave the same plate on the top the whole time, that plate will cool fastest and be most susceptible to condensation.


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OfflineSolipsis
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Re: How to prevent condensation in agar plates [Re: 50mmprime]
    #24553535 - 08/14/17 03:42 PM (6 years, 5 months ago)

What about covering a stack with something insulating like a teakettle cover, maybe even a warmed up one? What if you set them on something like a pizzastone that is cooled down. That kind of distribution of heat should discourage condensation I think?

I have tried putting a cup of warm water on top of the top dish of the shelf but unfortunately I don't have one that is large enough to cover it entirely and I am not exactly swimming in petri dishes especially not glass ones.

Yeah waiting until the agar is cooled down enough to hold is very important.

Really just thinking out loud here but I wonder what happens if you blowdry the top of a dish with condensation. Yeah you blow a lot of bacteria and stuff on there but if parafilmed so what?

Of course depends when you'd do that. Droplets of water just sitting on your agar isn't exactly cool either.

Good thread, interested to hear some thoughts from people more experienced than I. :smile:


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InvisibleEdiblesOnly
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Re: How to prevent condensation in agar plates [Re: Solipsis]
    #24553895 - 08/14/17 06:03 PM (6 years, 5 months ago)

This doesn't solve the condensation from building issue but it can help. If you have a flow hood u can open the Petri dishes and allow some air to move across the dish/lid. Now it's not good practice. But it does work on removing condensation


--------------------
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OfflineGr0wer
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Re: How to prevent condensation in agar plates [Re: EdiblesOnly]
    #24554369 - 08/14/17 08:34 PM (6 years, 5 months ago)

Pour cool around 120F, near gell stage. Put something hot on top like a coffee cup with hot water. A cheap IR thermometer will make testing temps easy https://www.amazon.com/dp/B00DMI632G?tag=myersmu-20


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OfflineMyc121416
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Re: How to prevent condensation in agar plates [Re: adadada]
    #25211547 - 05/17/18 10:45 PM (5 years, 8 months ago)

New to the site and I need a answer. I have agar dishes grown out but they are pinning inside the dish. Can I still ise them to spawn grain???


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OfflineSHROOMSISAY01M
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Re: How to prevent condensation in agar plates [Re: Gr0wer]
    #25211600 - 05/17/18 11:24 PM (5 years, 8 months ago)

Quote:

Gr0wer said:
Pour cool around 120F, near gell stage. Put something hot on top like a coffee cup with hot water. A cheap IR thermometer will make testing temps easy https://www.amazon.com/dp/B00DMI632G?tag=myersmu-20




The above is the best advice!!

I pour any leftover agar in a jar then microwave it to boiling and set it on top, it seems to stay hot longer than water. Make sure you have a vent on the lid like polyfill or a filter patch.


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OfflineQuadman
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Re: How to prevent condensation in agar plates [Re: Myc121416]
    #25211620 - 05/17/18 11:47 PM (5 years, 8 months ago)

Quote:

Myc121416 said:
New to the site and I need a answer. I have agar dishes grown out but they are pinning inside the dish. Can I still ise them to spawn grain???



Should be fine, a few pins wont hurt.


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OfflinebathingAPE
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Re: How to prevent condensation in agar plates [Re: adadada]
    #26535764 - 03/15/20 12:47 AM (3 years, 10 months ago)

I pour my plates at 105degreea with no issues as far as it gelling up on me before I get done with the pour. I've always wondered why people say 120? To me that's to hot, I always get condensation at that temp. My agar becomes un-pourable at around 98. So I've found that pouring between 110 and 100 has always yielded the best results.
I use a high quality laser thermometer to check temps.
I googled the Info as well, it says that agar gels up at 32-40 celcius.


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InvisibleBabuFrik
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Re: How to prevent condensation in agar plates [Re: bathingAPE]
    #26553353 - 03/23/20 03:51 PM (3 years, 10 months ago)

This is something I deal with in my lab all the time. If you use a pre-heated hot water bath to bring your temperature down to around 50-60°C right after it comes out of the pressure cooker, you can walk the solution down to a low temperature that is cool enough to hold in your hand and add temperature-sensitive antibiotics to it if that is something you do.

Reducing or outright preventing condensation on your plates is a cumulative effort but the steps involved are simple enough to follow for professionals and amateurs alike. Combine the lower solution temperature you get from the hot water bath with stacking your plates, a low RH in the room you are working in, and keeping the surrounding temperature the same throughout the polymerization process to help to reduce it. Lastly, if you have a flow hood or Bunsen burner and are confident in your equipment and method, you can crack the lids as the agar polymerizes to help release some of that local humidity caused by the cooling process.

I pour 20-40 plates about every two weeks or so in my lab and this method is very reliable. The only plates that see any condensation are those that sit on the top of the stacks and I can get rid of that by either cracking them in the hood for a little while, or by putting a 300 mL Erlenmeyer flask filled with hot water on the top. Please feel free to ask me if you have questions about this because I used to do this stuff as an amateur grower before I became a graduate student in a research laboratory and I am able to replicate the same process at home. :smile:


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Offlineseagu

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Re: How to prevent condensation in agar plates [Re: BabuFrik]
    #26553398 - 03/23/20 04:08 PM (3 years, 10 months ago)

so letting poured plates sit for a few minutes open in front of the flow hood after they gel up? Will this also get rid of the condensation from being stored in the fridge?


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InvisibleBabuFrik
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Re: How to prevent condensation in agar plates [Re: seagu]
    #26554999 - 03/24/20 11:58 AM (3 years, 9 months ago)

Quote:

seagu said:
so letting poured plates sit for a few minutes open in front of the flow hood after they gel up? Will this also get rid of the condensation from being stored in the fridge?




Let me make sure to double down on saying that this is a cumulative effort. There is no single solution unless you are lucky enough to have inadvertently accounted for some of the other variables. Pouring a cooler solution closer to its polymerization point, a working area with low RH, consistent temperature conditions throughout the process, and stacking plates as you pour should combine to solve most of the problem even into refrigeration. There might be a little fogging but that should abate normally. Heavy droplets do not usually form unless there is operator error, and that usually involves solution temperatures that are too hot or mixes that are too wet.

During the hot months when RH is in the 80s and climate control is not buffering it well, I pour a smaller number of plates unstacked and leave the lids cracked as they polymerize. The science behind this involves eliminating the local humidity in the dish released by the warm agar that would otherwise condense due to the higher humidity. When the lids are on there is only a small amount of air to buffer that extra moisture so it condenses, but leaving the lids cracked as the agar polymerizes allows the moist air to escape into the larger volume of air in the hood.

I have also poured and stacked as usual, then cracked the lids afterwards to try and evaporate the condensation like you are saying. That version takes longer when you have big droplets and the added time can cause the agar to dry out a little. That drying part is bad for me because I need precise concentrations for my research, but it is not a big concern for my gourmet workups since I am not collecting data. I won't recommend this version for pouring plates because the lid has to stay open for longer to get rid of more condensation. I see less condensation overall when I let the agar gel up while the lid is cracked. You can still do this to mitigate moisture that appears from cold storage, just let your plates warm up first to give the agar a chance to reabsorb some of it. If you have a big problem with condensation and you are lazy, you can always use 99% isopropanol to spray the lids and break surface tension of the water, forcing it to evaporate more quickly, but that is major corner cutting that requires some other precautions involving the spray bottle and filling it to be free of particulates.

For clarity, "cracked" means that the lid is partially off but not fully removed, something like 25-50% off. There is a non-zero risk associated with this technique, but if you are not an amateur, take all of these factors into account, and are confident in your equipment and technique, then you should trust in yourself not to be a vector. Everyone should be doing plate exposure testing on their flow hoods anyway to see if organisms are making it through the filter, so trust in the equipment to do its job and help overcome problems like these.


--------------------
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InvisibleSmartattack
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Re: How to prevent condensation in agar plates [Re: BabuFrik]
    #26557983 - 03/25/20 10:14 PM (3 years, 9 months ago)

I just put mine upside down on my table that happens to have a small space heater under it. You are basically running the physics in reverse, the condensate evaporates and re-condensates on the now cooler agar and if left overnight will just be re taken up by the agar. Works for me many times a week. All my top of stack plates condensate so they get this treatment.


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InvisibleBabuFrik
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Re: How to prevent condensation in agar plates [Re: Smartattack]
    #26558521 - 03/26/20 07:31 AM (3 years, 9 months ago)

Quote:

Smartattack said:
I just put mine upside down on my table that happens to have a small space heater under it. You are basically running the physics in reverse, the condensate evaporates and re-condensates on the now cooler agar and if left overnight will just be re taken up by the agar. Works for me many times a week. All my top of stack plates condensate so they get this treatment.




This reads like a good technique and the thermodynamics of this are sound. A light warming element underneath the plates should conduct heat into their systems. That gentle heating will support movement of air through the space in the plates' wells as they warm. My PI never passes up a chance to tell us that "heat goes out" of a system. To his credit, the localized humidity should be carried away with the exiting warm air into the cooler surroundings. I bet that this technique is also successful because most of the conduction is traveling through the top of lid and the condensation on it because the plates are upside down. I do something somewhat similar by stacking plates next to a Bunsen burner and rotating them over a time interval to help remove minor fogging around the edges when it is very humid.

This was interesting thank you for sharing your experience! Those top plates are the bane of people everywhere!


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If you are interested in trading mycology club tshirts, or want to help me buy one from your club, send a DM my way :banana:


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Offlinescarface710
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Re: How to prevent condensation in agar plates [Re: adadada]
    #26846436 - 07/26/20 04:42 PM (3 years, 5 months ago)

I know this is an old thread but the simplest way of avoiding condensation is by wrapping your plates correctly with parafilm. I have found that wrapping only one layer around the plate, minimizing overlapping layers has solved this issue. When I overlap to much of the parafilm a lot of the moisture gets trapped.


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InvisibleStipe-n CapMDiscord
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Re: How to prevent condensation in agar plates [Re: scarface710]
    #26846857 - 07/26/20 08:54 PM (3 years, 5 months ago)

Just leave your plates in front of the hood for an hour or so, or in your SAB unwrapped until they're gtg.
I personally let one side "defrost" like a windshield with the heat on in front of the hood for 30 mins then spin to the other side for another 30, then wrap like this:




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