0mar said:

nature always finds a way but im trying to be as proper as possible even if that yields headaches and massive disappointment... I'll get there some day hopefully

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tryptkaloids said:

I think that deserves its own thread.

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If the project is successful I will make one

Assuming my method and technique was good I expect in the 100 inoculation drops, let's be generous and say there are 50 isolated spores. Stamets reckons that only 1-5% of spores will germinate so averaging that at 3% and then halving it for using water agar gives me a 1.5% chance of any spore germinating. With 50 isolated spores that's a 75% chance on average of getting one.

I'll repeat it with proper germination plates in a few days.

covertjoy said:

The drops have set so I have been exploring one of the plates under the microscope. First drop I searched I didn't find any spores but on the second drop I found two stuck together.

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0mar said:

Bustaboo said:

Mine have a ton of condensation at first even when pouring at 117 degrees. It’s been clearing up pretty good after a day or 2.

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do you have them bagged up?

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plate.82 said:

Awesome!

I've been considering buying a microscope, there one you'd recommend?

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Mine is a second hand Motic B3 Series which I got for £200 on eBay and has brightfield, darkfield, phase contrast and polarisation modes. I do recommend a nice second hand one, microscopy enthusiasts seem to me to be people who look after their equipment and are honest, although that's a generalisation. The previous owner of this scope kept in touch and asks how I am getting on with it and gives me advice on how to use it effectively.

That said, for a new microscope the entry level AmScope microscopes have a good reputation. I used to have an equivalent one, the Swift 380T, which was good.

For looking into a petridish I use a very shallow pour and look through the underside. Using 4x and 10x objectives this works well (which combined with the 10x eyepiece gives 40x and 100x magnification). At 40x magnification (4x objective) the spores are just visible but difficult to differentiate from any debris, although I am able to cover a lot of surface area this way. At 100x (10x objective) the spores are more obvious.

To go up to 400x (which most recommend for mycology and which the photo in my previous post was taken at) you need a special objective if you want to look into petridishes. I added a new objective to my microscope which is a LWD (long working distance) 40x objective with a 0-2mm correction collar. The correction collar is very important as most objectives are configured to work with a 0.17mm cover slide which is not appropriate for a 1mm think petridish wall + agar. I got this objective second hand for £75 on eBay.



This is a single spore from the dilution inoculations. The image is somewhat crisper when looking through the scope, especially for 40x and 100x. The camera is where my microscope is weakest.

I actually found very few spores on the plates inoculated with the '2 spores per drop' solution although I was scanning quickly with the 40x magnification so probably missed some. The spore in the pictures here is from one of the plates with the '20 spores per drop' which I think is actually more like 1 spore per drop and the weaker solution more like 1 spore per 10 drops.

Not sure what the other structures are, they appear when I use solution from a test tube that has the dishwasher rinse aid added. Could be to do with the rinse aid or could be to do with the fact that I pressure cooked these plastic (polypropylene) test tubes with distilled water in them before making the solutions. Actually I find that these structures are useful for finding where the drops are for searching.

covertjoy said:

For looking into a petridish I use a very shallow pour and look through the underside. Using 4x and 10x objectives this works well (which combined with the 10x eyepiece gives 40x and 100x magnification). At 40x magnification (4x objective) the spores are just visible but difficult to differentiate from any debris, although I am able to cover a lot of surface area this way. At 100x (10x oTo go up to 400x (which most recommend for mycology and which the photo in my previous post was taken at) you need a special objective if you want to look into petridishes.

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Screwup said:

Look at this nerd actually adhering to the whole “spores are for microscopy use only”.

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tittlez said:

Can you also see bacterial contamination at x400 magnification?

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I looked at a bacteria colony at 400x and it is super fascinating. It created a very beautiful geometrical pattern. I am not sure if I was looking at the actual bacteria or the affect they have on the media. I don't have photos of that but next time I get a chance I will get some. I believe that if you know what you are looking for you would be able to identify bacterial contamination at 400x. I don't have that experience personally but it is something I would like to learn. I think that different bacteria would present differently.

Edit:

Turns out I still have that plate.



This is a bacterial colony. The green dash lines I added denotes the leading edge, which is more visible through the scope.

The geometric pattern is not organised like it was when I first looked at this. It might be because the culture moved when I picked up the plate.

Here is an image from the internet: