Pics of Yeast under my new microscope

[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

 

[passedpawn]

Here's how my scope looks with the little OEM video camera on it. The camera is mounted onto the eyepiece tube. There is no lens on the camera. The camera looks like the following. It's held onto the scope by a highly sophisticated arrangement of rubber bands.

 

[smokinghole]

Violet is better because it's a more pure stain and it has a lowered incidence of false viability. In the Yeast Performance book I linked that no longer has a preview, they showed that methylene blue showed viability up to 15% or greater on heat killed cells. Methylene violet showed near 0% viability.

 

[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.

Brewing with this yeast was officially a mistake (and learning experience). I kegged it and left the carboy with yeast sitting on my patio (I was thinking about harvesting the yeast). Uh, no. This, after 2 days:

The beer looks good, but there is a slight funk there. It's a dry stout. Just barely tolerable, but I'm dumping this weekend.

 

[SGFBeerBuzz]

Just a slight funk? Sounds tasty. I like a sour stout.

 

[passedpawn]

Just a slight funk? Sounds tasty. I like a sour stout.

Actually the stout turned out OK. I thought it would get worse, but it didn't and is probably not completely out of style for a dry stout.

 

[jagec]

Actually the stout turned out OK. I thought it would get worse, but it didn't and is probably not completely out of style for a dry stout.

The great thing about beer is that it almost always gets better, not worse, if you just forget about it for a while!

 

[COLObrewer]

Actually the stout turned out OK. I thought it would get worse, but it didn't and is probably not completely out of style for a dry stout.

Dood! I'm glad you ended up following the #1 rule, never dump your beer.

I was wondering if you saved any of your juniper yeast and looked at it? I haven't searched through this thread though.

 

[passedpawn]

Dood! I'm glad you ended up following the #1 rule, never dump your beer.

I was wondering if you saved any of your juniper yeast and looked at it? I haven't searched through this thread though.

No, I didn't. Would have been interesting though. If you ever want me to get some pics of yours, PM me and I'll give you an address.

 

[H-ost]

Anyone have anymore pics of yeast to add? These are fun to look at.

 

[passedpawn]

Anyone have anymore pics of yeast to add? These are fun to look at.

I've got loads; maybe I'll get some more up here.

The Brett Project is a neat site put up by some dude working on his masters thesis. Check it out. Some pics of yeast, smears, etc.

http://www.brettanomycesproject.com/2010/06/brettanomyces-yeast-cell-images/

 

[14thstreet]

Not much information here but maybe a few of you would like to see naked yeast

Picture 1 is a third or fourth generation washed sample of Wyeast 3068. Trypan blue staining to check viability, which was about 10%, after 8 months in the refrigerator. Note some sort of coccus strain, perhaps our friend Pediococcus (diacetyl, sourness, etc). I won't be culturing this up for my upcoming Oktoberfest party.

Picture 2 is a harvest from Lakefront Brewery's Wisconsinite from earlier this summer. Looks very different from 3068 and is much cleaner. Viability was determined from a different sample and was 6%. I might try stepping this culture up to see how it goes.

 

[passedpawn]

Not much information here but maybe a few of you would like to see naked yeast

I love naked yeast! What type of scope / objective? How are you taking pics (trinoc?)

Alright, I'm gonna do some viability stuff and get some more pics up here!

 

[passedpawn]

eyeMicroscope iPhone app assists picture taking on your microscope. Looks sorta neat for anyone who is taking cell phone pics through the eyepiece. All the app does is store the scale bar next to the pic, which is pretty neat. Worth $3? You be the judge.

[edit] actually, I was just experimenting with taking iPhone pics through my eyepiece and it sux. Unless you've figure out how to do this easily, don't bother with the app.

 

[diS]

Alright, I'm gonna do some viability stuff and get some more pics up here!

Please do, when I see this thread bolded I get excited and hope we"ll see our buddies..

Also, it would be great if some of you advanced guys/girls make some viability testing, like w/yeast frozen in glycerin solution...

 

[14thstreet]

I love naked yeast! What type of scope / objective? How are you taking pics (trinoc?)

Alright, I'm gonna do some viability stuff and get some more pics up here!

This was done at work on an inverted Nikon microscope with plastic slides, 400-600X. It wasn't an ideal setup because I had other work to do and I don't want to waste resources. Sorry I'm not more specific but this isn't a do-it-at-home type microscope.

Regarding glycerin stock viability, I could do something like that. I'd have to come up with an experiment that would be practical. I think with 6+ months in the fridge with a multi-repitched wheat strain I'm pushing my luck with sanitation and other qualities.

 

[passedpawn]

COLOBrewer sent me some of the yeast he harvested off juniper berries found in the wilds of his state. Here's some pics. I've got video I'll add to this later of me staining it with violet and blue. Looks like normal Saccharomyces to me.

More later... hot date to attend to.

Watch as I add a drop of violet stain to the hemocytometer. The ones that go dark are dead.




Here it is, plated and after 2 days of growth.

 

[COLObrewer]

Very interesting sir, looks like you are learning alot about yeast, I can hardly wait for your morphology experimentings.

 

[passedpawn]

Plated Berliner Weisse yeast. There is Lactobacillus Delbrueckii in there and Saccharomyces (S-05) too. I looked at the cells before I plated it and it looked just like sacc, but these long stringy things probably are bacterial growth. No idea what I'm saying here, but it sure sounds official.

Here's the plate on the scope stage. I accidentally stuck the 100x objective into it when I was pushing the lighting condenser up from below. Plate is contaminated, need to clean objective. Meh.

My scope:

Stuff (lacto?) growing across the plate

 

[passedpawn]

I'm making a 15 minute video of this bacteria. It's growing pretty fast. I'll speed it up into a 30 second video and we'll all get to see the growth in fast mo! Stay tuned. (Is there anybody tuned?)

 

[passedpawn]

Ok, this is sorta cool. I recorded 22 minutes of that bacteria growing across the slide. I then dropped that video into Pinnacle Studio and compressed it into 2 seconds, then uploaded to you tube.