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OfflineStrata
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Registered: 03/23/13
Posts: 16
Loc: UK
Last seen: 11 years, 28 days
Is a spore likely to germinate under a coverslip?
    #18013673 - 03/26/13 12:13 PM (11 years, 1 month ago)

Just wondering, I'd like to get some really nice 'scope images of fungal growth - including timelapse, trouble is a really nice 'scopes tend to be immersion ( all ours are anyway - especially the ones with robotic stages ) .

I think the easiest way would be to dot out diluted spore mixture onto a low melt agar ( liquid above 25C )then coverslip it immediately.

But - a coverslip isn't the most oxygen permeable thing in the world and will mycelium cope along the agar-glass interface? -  since I can't image through an air gap. With more than one spore on a slide I should be able to get images of fungi getting their end away.

An alternative would be to drill some slides, permanently fix a coverslip then load with a thin layer of agar through the hole before covering in permeable tape - then attempt to image through the agar but that would be a lot of work for something I want to do on the qt after hours.

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OfflineNobitte
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Registered: 02/20/08
Posts: 493
Loc: Biosphere
Last seen: 6 years, 7 months
Re: Is a spore likely to germinate under a coverslip? [Re: Strata]
    #18015657 - 03/26/13 06:44 PM (11 years, 1 month ago)

This sounds great, I would say that it is worth a try, I've had spores germinate in sealed solutions a bunch of times.

It shouldn't have a problem with being in close proximity to the glass, although it might have trouble with lack of GE, although if its just a cover slip, I would imagine that diffusion from the sides alone would suffice.

it would depend on the species as well, as to how tolerant they are of anaerobic sealed conditions like that.

There are a few time lapses on youtube, perhaps you could contact the person who made them and ask them for tips.

Please keep me/us posted xx

Nobitte


--------------------
First we must learn...

Then... WE CAN TEACH :weirdeyes:

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OfflineStrata
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Registered: 03/23/13
Posts: 16
Loc: UK
Last seen: 11 years, 28 days
Re: Is a spore likely to germinate under a coverslip? [Re: Nobitte]
    #18017152 - 03/27/13 01:14 AM (11 years, 1 month ago)

I'll start with pleurotus species as a convenient and quick growing choice then maybe try with P.stipticus ( if it ever fruits for me ) - unless I come across something faster germinating. I took the liberty of setting up a remote desktop for the microscope controller so I can check progress over the weekend :smile:

This also strikes me as a good way to get monokaryotic mycelium and a route to making my own oyster hybrids.

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OfflineNobitte
Student


Registered: 02/20/08
Posts: 493
Loc: Biosphere
Last seen: 6 years, 7 months
Re: Is a spore likely to germinate under a coverslip? [Re: Strata]
    #18023608 - 03/28/13 12:08 PM (11 years, 1 month ago)

Good luck, I am rather interested to see what comes of it, it seems like it could be quite effective indeed.

Also, if you cant find anything similar you might want to stay tight lipped about your creation and get a greedy patent, but feel free to message me about it =3.


--------------------
First we must learn...

Then... WE CAN TEACH :weirdeyes:

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Offlinesmegzilla69
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Registered: 09/02/12
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Re: Is a spore likely to germinate under a coverslip? [Re: Nobitte]
    #18034501 - 03/30/13 07:34 PM (11 years, 1 month ago)

:popcorn:


--------------------

EVERYTHING I SAY IS A LIE!!! N ALL MY PICS R STOLEN
WORTH EVERY PENNY!!!!watch it pay for it self in # of high yield flushes!
SO DOPE
"The true sign of intelligence is not knowledge but imagination. "  Albert.

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OfflineStrata
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Registered: 03/23/13
Posts: 16
Loc: UK
Last seen: 11 years, 28 days
Re: Is a spore likely to germinate under a coverslip? [Re: smegzilla69]
    #18041220 - 04/01/13 05:10 AM (11 years, 1 month ago)

Well I'm buggering of to Cuba for a couple of weeks,  and doing a little spore hunting whilst I'm there so I'll set up when I get home.  Since I have no fruiting bodies at the moment I'll probably start with the old bisporus as the easiest option for getting a print.  Try it on the home microscope before moving onto the hulking great delta at work.

Anyone dare to guess how long it takes to get microscopically visible germination? It'd be nice to get the frame  interval right first time

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OfflineNobitte
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Registered: 02/20/08
Posts: 493
Loc: Biosphere
Last seen: 6 years, 7 months
Re: Is a spore likely to germinate under a coverslip? [Re: Strata]
    #18043324 - 04/01/13 06:07 PM (11 years, 1 month ago)

Depending on the media, and don't quote me on this, but the monokaryons should be visible within the first few days, although this is likelt to be variable with both species and conditions such as substrate content relative to requirements/triggers, GE, incident light and temperature.

Hopefully someone else can fill you in on that one, I have little experience with actually watching spores germinate.

Im just going on how long it takes me to see dikaryons with the naked eye(on agar or grain) and assuming that the process probably starts several days before even the first white spot is visible, meaning that in ideal conditions it might occur within hours of them coming into contact with the substrate.

Itd be interesting to come up with a vague timeline as to their germination and eventual plasmogamy. It might be worth looking at some conidia at some point as well.

I wonder what factors stimulate germination, id do some reading but im in the midst of a very busy day, im sure you understand.

If I ever decide to steal this idea or something related to get a grant/research, ill tell you. Also, if you need spores to play around with, I have a few spares in my collection that I could send your way to have a look/play with.

Have fun in Cuba ^^

Nobitte xxx


--------------------
First we must learn...

Then... WE CAN TEACH :weirdeyes:

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