Yeast cells under microscope

[JohnB]

Greetings everyone! I am posting some photos I took under a microscope at a dilution 1:4000.

I used methylene blue to stain the cells and although there are a lot dead ones, there are still some viable ones and have calculated them at 800.000.000/ml . I pitched about 350ml in total and the age of it was 2 1/2 months in the fridge at 4C.

I also see something that looks like tissue or fiber that is also stained blue and wanted your opinion on whether or not this is some bacteria.

I pitched this yeast in a 1.060 IPA 0.822 IBU/SG

Fermentation is going and I guess if the yeast had been infected I may get away with it because I used it in an I.P.A.?

 

[passedpawn]

I only see yeast cells - no bacteria. They also all look dead

You would benefit from more magnification (though, perhaps the photo is the problem - looks like a cell phone shot through the eyepiece tube). Is that 100x?

It's pretty normal to see a few random bacteria cell in your starter. They are everywhere. Look for rod shapes, much thinner than the oblong/round yest cells.

 

[passedpawn]

I see a giant hydra in that first photo. Kraken! He's gonna take down the whole ship - beware

 

[JohnB]

Thank you, Yes it is a photo taken by my cell magnification was 200X and 400X

The weird think is that I took another 4 samples without staining them and only yeast cells were visible and nothing else .

Then I stained a fifth sample and again there was nothing else but cells .

I was thinking that something might be in the methylene blue solution, but like I said nothing showed up in the fifth sample .

P.S. Yesterday I got my hands on a crystal violet solution and after I stained a different sample,OMG there were hundreds of tiny "crystals" everywhere at 400x. I shook it as well as possible but to no avail.

 

[passedpawn]

Thank you, Yes it is a photo taken by my cell magnification was 200X and 400X

The weird think is that I took another 4 samples without staining them and only yeast cells were visible and nothing else .

Then I stained a fifth sample and again there was nothing else but cells .

I was thinking that something might be in the methylene blue solution, but like I said nothing showed up in the fifth sample .

P.S. Yesterday I got my hands on a crystal violet solution and after I stained a different sample,OMG there were hundreds of tiny "crystals" everywhere at 400x. I shook it as well as possible but to no avail.

I used to use the violet. I did not have any crystals. Did you get it in crystalline form, or was it liquid already?

 

[JohnB]

I used to use the violet. I did not have any crystals. Did you get it in crystalline form, or was it liquid already?

It's in liquid form. I will try to upload a photo...Please see attached photos

 

[passedpawn]

It's in liquid form. I will try to upload a photo...Please see attached photos

That's the one on Amazon. I got mine from White Labs. I need more, maybe I'll try that Amazon one.

I wonder why the crystals didn't dissolve?

 

[JohnB]

Hmmm, I thought to use this time less crystal violet, than a final solution of 1:1 as I have read pretty much everywhere, and as you will see there are no crystals! I used 2 drops in a 40ml de-ionized water .

P.S. I think I am getting better with the photos

 

[passedpawn]

Your cells look like they are huge - not sure why that is. Here's a pic I took where you can see those squares - note that those squares are 20 µm on a side. That means that the yeast are about 3 or 4 µm long. These lines are pretty standard on these counters, so I'm not sure why the cells on your's look like they fill the square.

 

[passedpawn]

Hmmm, I thought to use this time less crystal violet, than a final solution of 1:1 as I have read pretty much everywhere, and as you will see there are no crystals! I used 2 drops in a 40ml de-ionized water .

P.S. I think I am getting better with the photos

I don't think the things in that last picture are yeast cells. I'm no biologist, but they look much too large to be beer yeast cells.

 

[JohnB]

No wait...the last picture has no yeast cells, just de-ionized water and 2 drops of crystal violet. It was just my point that by using only 2 drops in a 40ml de-ionized water I didn't see any crystals as when I used a 1:1 (1ml to 1ml) in the final solution (yeast - de-ionized and crystal violet.) Wait a second now that I am thinking about it does "crystal" violet mean that there are crystals in it and perhaps in larger concentration as 1:1 you see them??? Definitely I will try a 1:1 again tomorrow

The first pictures on the the thread have yeast cells

 

[JohnB]

That's the one on Amazon. I got mine from White Labs. I need more, maybe I'll try that Amazon one.

I wonder why the crystals didn't dissolve?

By the way, yes I got it from Amazon.