Pics of Yeast under my new microscope

[rhamilton]

That paper considers the ester development of the finished beer, not yeast viability. Still seems like an interesting test.

Recommended addition is 1mg olive oil per 25B cells. For a smack pack that's 4 mg of olive oil. How does one measure that? I'd probably go to 1 gram.

Doesn't olive oil float? Does it need to get boiled with the starter wort?

Yea you'd need a micro-liter pipette. The oil would float but between a stir plate and natural yeast currents, it should be readily available to them. I don't know about the boiling part -- I assume olive oil is fairly clean since it's stored at room temp and it doesn't spoil.

Most people who use EVOO (props to Miss Ray ) use a drop off of a fine needle since it's such a small amount.

The reason I ask is the research done by New Belgium related to it. They apparently stopped using it due to shelf life issues but my beers don't last that long anyways. I was debating an aeration setup when I came across the EVOO research and I was intrigued but I felt the research was lacking in terms of yeast propagation. If it greatly improves yeast growth, it would be a great addition to making starters.

 

[passedpawn]

I went ahead and added olive oil to a starter tonight. Altbier yeast for a dry stout I'm making in a week. What the heck.

 

[kcpup]

I went ahead and added olive oil to a starter tonight. Altbier yeast for a dry stout I'm making in a week. What the heck.

I'm interested to see how the test turns out!

 

[rhamilton]

I'm interested to see how the test turns out!

+1 -- how did it turn out with the EVOO?

 

[passedpawn]

+1 -- how did it turn out with the EVOO?

I grew some starters, but then froze them (and all my beer) in my keezer. Beer is OK, yeast probably not so good. I need to start over.

I will say, though, that the starter was raging without any path for O2 ingress.

I'll repeat the experiment this week.

 

[passedpawn]

Dregs
Dregs from Smuttynose Farmhouse Saison. I was expecting to see some Brett here. I think there might be some brett cells there... or maybe all of them are. No idea. I've tried to compare to pics at some other sites. I think brett should be longer, like the cell in the middle of the pic, and the Sacc is more round or egg-shaped.

Stain
FYI, violet stain added to look for dead cells. Instead of adding the stain to my mixing vessel (shot glass!), I tried adding a drop right to the slide after the sample was loaded and it worked great! That makes it super easy. Gonna try this with the blue stain too.

Video/Snapshots
I have a small OEM camera module mounted to the eyepiece tube. It's made by a company called IDS. I get a video stream, and from that I can take snapshots. There are a million video settings that I'm playing with to get decent pictures (and video). Mostly, I play around with the white balance to make the field look gray or white. It's a work in progress.

 

[dstar26t]

FYI, violet stain added to look for dead cells. Instead of adding the stain to my mixing vessel (shot glass!), I tried adding a drop right to the slide after the sample was loaded and it worked great! That makes it super easy. Gonna try this with the blue stain too.

Only problem with that is you don't know your dilution factor anymore and can't determine viability.

 

[passedpawn]

Only problem with that is you don't know your dilution factor anymore and can't determine viability.

Yea, that's true. Darn it.

 

[passedpawn]

I accidentally froze this yeast, so I grew a starter from the frozen stuff - that's why the violet stain is there - looking to see how many I killed.

But man, look at this crazy yeast. This is the yeast I brought back from Fuchschen Brewery in Dusseldorf Germany. Those "long-skinnies" have to be wild yeast, right? They use open fermenters at the brewery, so I suppose it should be a surprise, but it is anyway.

 

[jagec]

For anyone who is interested: Some Wyeast 3787 under high magnification DIC, no stain.

 

[passedpawn]

For anyone who is interested: Some Wyeast 3787 under high magnification DIC, no stain.

Very cool. DIC gives amazing contrast. Do the images always look dark like that? That looks like a lot of magnfication... ~ 1000x. What type of scope is that? Was the objective an oil immersion?

(At this moment I'm looking at the bacteria in my own spit, plus some alarmingly large single-cell things that I have no name for; in truth, I can't name anything I see under a scope except for yeast cells ).

 

[jagec]

Very cool. DIC gives amazing contrast. Do the images always look dark like that? That looks like a lot of magnfication... ~ 1000x. What type of scope is that? Was the objective an oil immersion?

(At this moment I'm looking at the bacteria in my own spit, plus some alarmingly large single-cell things that I have no name for; in truth, I can't name anything I see under a scope except for yeast cells ).

You actually vary the orientation of the Nomarski prism to control the amount of contrast. If it's parallel to the polarizers, you get maximum contrast, and the image is almost completely dark except for the edges of the cells and some of the internal structures, which are fiendishly white. If you deoptimize it, the image looks like bright field, and you lose the ability to see some of the fine features. Generally it's best to use an orientation that highlights the features without completely drowning out the rest of the field.

The scope is an Olympus IX71, and the objective is a 60x oil immersion with a 1.5x optivar, and I think a 10x eyepiece. I forgot to put a scale bar on that picture, but the big cells are about 10 microns along the long axis.

 

[passedpawn]

You actually vary the orientation of the Nomarski prism to control the amount of contrast. If it's parallel to the polarizers, you get maximum contrast, and the image is almost completely dark except for the edges of the cells and some of the internal structures, which are fiendishly white. If you deoptimize it, the image looks like bright field, and you lose the ability to see some of the fine features. Generally it's best to use an orientation that highlights the features without completely drowning out the rest of the field.

The scope is an Olympus IX71, and the objective is a 60x oil immersion with a 1.5x optivar, and I think a 10x eyepiece. I forgot to put a scale bar on that picture, but the big cells are about 10 microns along the long axis.

Is that inverted? How do you use an oil immersion when the objective is facing up?

 

[jagec]

Is that inverted? How do you use an oil immersion when the objective is facing up?

Yes, I don't think anyone is using non-inverted microscopes in research labs these days.

You just put a droplet of oil on the objective and put the coverslip with your sample on it on top. The oil is thick enough to stay in place. If you move your sample around a bunch, the bottom of the coverslip gets oil smeared all over it, but that's not really a problem.

 

[pabloj13]

Yes, I don't think anyone is using non-inverted microscopes in research labs these days.

You just put a droplet of oil on the objective and put the coverslip with your sample on it on top. The oil is thick enough to stay in place. If you move your sample around a bunch, the bottom of the coverslip gets oil smeared all over it, but that's not really a problem.

That's not really true. There are a lot of applications where an upright microscope is far superior. For example, making electrophysiological recordings from brain slices.

 

[cmybeer]

So I just aquired a pretty sweet vintage microscope. It's a baush and lomb and my best guess is that it is from the 1930-50 range. The eye piece is 10x magnification while the three lenses are 10x, 43x and a 97x oil immersion lens so there is more than enough magnification power.

The problem (I think) is that there is no light source or condenser of any kind. I know nothing about microscopes but think that doing all this stuff would be an interesting way to up my interest in homebrewing even more. Can I just stick a light under there, or get a mirror. Worst case scenario I think I could sell this for a lot of money and buy a newer one that has everything included.

 

[passedpawn]

Yes, I don't think anyone is using non-inverted microscopes in research labs these days.

You just put a droplet of oil on the objective and put the coverslip with your sample on it on top. The oil is thick enough to stay in place. If you move your sample around a bunch, the bottom of the coverslip gets oil smeared all over it, but that's not really a problem.

Sooo, I guess this isn't a scope in your home.

Those lab scopes from Olympus are pricey. A bit pricey for the home hobbyist looking for hydra in pondwater. I'll stick with my Oly CH2 ( until one of my investments does something )

 

[pabloj13]

So I just aquired a pretty sweet vintage microscope. It's a baush and lomb and my best guess is that it is from the 1930-50 range. The eye piece is 10x magnification while the three lenses are 10x, 43x and a 97x oil immersion lens so there is more than enough magnification power.

The problem (I think) is that there is no light source or condenser of any kind. I know nothing about microscopes but think that doing all this stuff would be an interesting way to up my interest in homebrewing even more. Can I just stick a light under there, or get a mirror. Worst case scenario I think I could sell this for a lot of money and buy a newer one that has everything included.

You'd be able to shove a light source under there, but you're only going to be able to use brightfield illumination with that setup. But you should be able to see them.

 

[passedpawn]

You'd be able to shove a light source under there, but you're only going to be able to use brightfield illumination with that setup. But you should be able to see them.

At high magnifications he'll need a lot of light, something focused. But it could be done.

 

[william_shakes_beer]

I'd be interested in getting enough magnification to verify that there is far more yeast than bacteria in my harvested storage strains. What kind of magnification would I need to do that?

 

[passedpawn]

I'd be interested in getting enough magnification to verify that there is far more yeast than bacteria in my harvested storage strains. What kind of magnification would I need to do that?

400x is best, but you could probably differentiate between yeast and bacteria with 200x.

Most microscopes have 10x eyepieces, so you'd need that then 20x or 40x objective lens.

Further, make sure it has a light source with condensor under the stage (no mirrors), X/Y mechanical adjustment so you can move the slide around without your fingers. Binocular is VERY nice compared to monocular. Major brands are Olympus, Leica, Zeiss, and Nikon. Some other brands (i.e., AO or American Optics) might be OK, might not. I have no experience with the very cheap ones that connect to a computer.

 

[jagec]

That's not really true. There are a lot of applications where an upright microscope is far superior. For example, making electrophysiological recordings from brain slices.

I guess my cell bio bias is showing.

 

[pabloj13]

I guess my cell bio bias is showing.

No worries. I'm in a cell bio department too.

 

[cmybeer]

At high magnifications he'll need a lot of light, something focused. But it could be done.

I was just messing around with my camping headlight which is pretty bright and I pulled out a piece of hair to try to see it in focus. I got it on the 10x but seeing that I don't have any slides or anything I was having troubling getting the sample to stay in place for the 43x. It seemed fairly bright. I'll mess around with it a bit more to see if I can get it in focus. Maybe I could pull this off.

If not I was doing some pursuing for used parts online and found pretty much the same scope (albeit with a lot more accessories and a fancy wooden box) on ebay for $700.

 

[passedpawn]

I was just messing around with my camping headlight which is pretty bright and I pulled out a piece of hair to try to see it in focus. I got it on the 10x but seeing that I don't have any slides or anything I was having troubling getting the sample to stay in place for the 43x. It seemed fairly bright. I'll mess around with it a bit more to see if I can get it in focus. Maybe I could pull this off.

If not I was doing some pursuing for used parts online and found pretty much the same scope (albeit with a lot more accessories and a fancy wooden box) on ebay for $700.

If you could sell yours for that, you'd be able to get a great scope, hemocytometer, stains, slides, pipettes, and you'd soon be looking at your own spit.

 

[cmybeer]

Yeah spit! That one was in much more complete condition but even if I could get half that. We'll see, my girlfriend got it from her parents so I don't know if I should sell it.

 

[passedpawn]

No worries. I'm in a cell bio department too.

I'm in my den with a cold one next to my scope

 

[passedpawn]

I changed video cameras on my scope:

Previous:

• IDS UI-1556LE-C-HQ
• 1/3" CMOS sensor
• 1600x1200 pixels 18 fps

New:

• IDS UI-1466LE-C-HQ
• 1/2" CMOS sensor
• 2048x1536 pixels 11 fps

I did side-by-side tests and the new camera is markedly better. Pixel noise and resolution improvements are noticeable. Woohoo.

Here's some Wyeast 2565 Kolsch yeast right out of the starter with methylene Blue stain. Cells are budding all over the place.

 

[cmybeer]

Very nice! I just put in the order for my hemocytometer the other day. Got some slides and cover slips last week to give it a test run and to make sure every thing was visible clearly enough. It was so I bought stuff to do some counting. I'm excited.

I still need some stain. Would you just recommend the methylene blue?

 

[passedpawn]

Very nice! I just put in the order for my hemocytometer the other day. Got some slides and cover slips last week to give it a test run and to make sure every thing was visible clearly enough. It was so I bought stuff to do some counting. I'm excited.

I still need some stain. Would you just recommend the methylene blue?

Now that I've used both methylene blue and the methylene violet, I like the violet better. The stain is easier to see. I don't have much more reason than that. I think there are better reasons to use it, but I don't have enough data to state anything conclusively.

Check your local fish stores. The ones that serve the saltwater and reef aquaria hobbyists might have methylene blue. It's used to treat the water in some way.

If you can't find a stain locally, just buy from White Labs. Get both, they are really cheap. Let me know how it turns out for you.l