Pics of Yeast under my new microscope

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Well.....I am intrigued. I dont unnerstand the need for a scope but I like anything techy anyway. What is the purpose of using a scope. To visually identify different strains or to verify live cells percentages? How would it improve my process and what is the cost versus return on investment. I know a couple of jacked up batches could pay for that equipment, but I just need help with the need to employ that kind of gear. TIA for any answers.
Bob
 
bierhaus15 said:
Can someone give me a short rundown on the cost and type of equipment I would need to get into this? I have a very old scope that I haven't touched since HS, but I doubt it would work for this. I know my local college sells used lab equipment a few times a year.
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If you can do 400x and see clearly you're good. I keep watching craigslist for a good scope for myself at home. Then you'll need a hemocytometer which aren't super expensive I think they can be had for less than $100. Then lastly you'll need some dye, methylene blue is the old standard, but many places are moving to more purified and reliable dyes like methylene violet (3rax). You can buy the solutions from white labs or you can look up how to make your own.
 
smokinghole said:
If you can do 400x and see clearly you're good. I keep watching craigslist for a good scope for myself at home. Then you'll need a hemocytometer which aren't super expensive I think they can be had for less than $100. Then lastly you'll need some dye, methylene blue is the old standard, but many places are moving to more purified and reliable dyes like methylene violet (3rax). You can buy the solutions from white labs or you can look up how to make your own.
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Hemocytometer on ebay was about $30.
 
passedpawn said:
M

Based on the dilution and the number of cells there, I get 82.5 billion in the pouch. I only counted one cell area, so lots of possible error there. My dilution is using disposable pipettes, could be quite a bit of error there too. But the result is very close to what Wyeast states. BTW, I've measured about 150ml from a smack pack after the internal nutrient pouch is broken and mixed in. Here's the spreadsheet I made to do my counts... note the Total Cells in Flask (which in this case is the 150ml pouch).


Yeast_Counting_Spreadsheet_2.jpg
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I like the calculation that you made. What is the application are you using to make it?

I had forgotten that I have a microscope and hemocytometer in storage from some of my wife's hobbies. This thread reminded me of that, so I got them out and want to start making actual counts of my starters.
 
Rbeckett said:
What is the purpose of using a scope. To visually identify different strains or to verify live cells percentages?
Click to expand...

Hard to visually tell the difference between strains of yeast from what I've seen. They don't look identical, but telling the difference is probably not the point of this. This is mostly to verify cell counts so you can pitch the correct amount of yeast

Rbeckett said:
How would it improve my process and what is the cost versus return on investment.
Click to expand...

Honestly, if you're already making yeast starters and paying attention to yeast health (oxygenation, etc). I doubt you'd see a huge improvement from this. Certainly not a good return on investment as this equipment is probably about the cost of 20 batches or more. This is a hobby, getting into the microbiology side of it is just another way to have fun with it.
 
So I didn't see much activity in my starter. I wanted to take a count to see if I had growth. I had an infection. I saw bad bacteria swimming around so I ditched it.
 
Well I got a microscope and have been checking out all my yeast. I had an infected batch about two years ago, I put the yeast under the microscope and saw the bacteria or wild yeast. I do not know what it is. I tried to get a good video of it, but my camera keeps zooming in and out on its own trying to focus.

Sorry for the bad quality, you might want to mute it as well I didn't realize I was listening to music at the time.

http://www.youtube.com/watch?v=qjyqMZh_e3s&feature=youtu.be

At 44 seconds I catch a long looking thing. There are tons of them all mixed in with yeast, wish you guys could see what I am seeing that is really crap quality.
 
GodsStepBrother said:
Well I got a microscope and have been checking out all my yeast. I had an infected batch about two years ago, I put the yeast under the microscope and saw the bacteria or wild yeast. I do not know what it is. I tried to get a good video of it, but my camera keeps zooming in and out on its own trying to focus.

Sorry for the bad quality, you might want to mute it as well I didn't realize I was listening to music at the time.

http://www.youtube.com/watch?v=qjyqMZh_e3s&feature=youtu.be

At 44 seconds I catch a long looking thing. There are tons of them all mixed in with yeast, wish you guys could see what I am seeing that is really crap quality.
Click to expand...

Yea, I see that chain thing there. Doesn't look good!
 
bruin_ale said:
Looks like some type of Brett probably. Bacteria would appear much smaller than that.
Take a look at these pics, some of them seem to resemble the rod/tube shaped cells from your video.

http://brettanomyces.wordpress.com/
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Yep that is it, I have had this beer for 2 years and it has never gotten better. Funny to actually see them though, I am going to try and introduce some pure alcohol and see how long it takes for them to die, and then I will try star san. Should be fun:ban:
 
Bell's Oberon dregs (puportedly a defunct Wit strain). With Alkaline Methylene Violet stain.

I probably used too much stain, but it looks like there's only 1 viable cell there! I put an arrow next to it (while I was finishing that Oberon).

stain_-_violet_-_oberon.jpg
 
I'm not sure about using the AMV stain but the way I do the MB stain is this:

Combine 1ml stain with 1ml yeast slurry
Let stand for 60 seconds after mixing well
Perform serial dilution
View in scope

My method slightly varies from the normal quoted method. I have done it both ways and I get very similar results, but my way is just easier to see the stained cells. When you add the 1ml of dye to the final dilution you're staring at solid blue with cells and depending on the scope it can be tough to differentiate.

Now with AMV the beauty appears to be that if it is pink at all they're non viable. You can clearly see some of them are lysed or just in general screwed up. Others have the dye slightly pooled up, I would be inclined to count them as dormant not dead. I suspect that budding cells will not metabolize the dye out either so they will pick up dye. You just have to get a feel for things as you become more familiar with the process.
 
martinworswick said:
i'm shopping around for the bits required for my own lab,it's taking a while due to my meagre budget.

can anyone offer any opinions regarding the suitability of the following items?

http://www.trademe.co.nz/Browse/Listing.aspx?id=463688327

http://www.trademe.co.nz/Browse/Listing.aspx?id=465059462

cheers
martin
Click to expand...

hemocytometer looks fine. Nice box.

Scope illumination might be a problem. That's a reflector at the bottom. Hard to get enough light with that at high magnification. I'm no expert though.
 
passedpawn said:
hemocytometer looks fine. Nice box.

Scope illumination might be a problem. That's a reflector at the bottom. Hard to get enough light with that at high magnification. I'm no expert though.
Click to expand...

cheers,
i'll keep my eye on the auction for the hemocytometer and i'll do a bit mor research into the scope. maybe if i rig a bright light straight in front of the illuminator? or replace it with a halogen or similar? mmm i wonder.

most of the decent scopes are out of my budget over here, and freight costs kill the savings from shipping one in!
 
So for those of you that have them or those of you who have experience with them, what qualities should one look for in a microscope? I mean, there are ones on amazon for about $35 which seems like a bad idea to me. And then there are the student recommended ones that are around $125.

I'm thinking of picking up the Yeast book, is this kind of stuff outlined in there? I would love to nerd out on yeast, I really enjoyed this stuff during highschool and actually thought I wanted to study biology in college until I realized how much harder it was and how much math and chemistry I would need.
 
OP -- would you consider doing a olive oil experiment with a few starters? I'd be curious to see how it effected actual yeast viability / counts.
 
rhamilton said:
OP -- would you consider doing a olive oil experiment with a few starters? I'd be curious to see how it effected actual yeast viability / counts.
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You're going to have to spell it out for me -- I don't know what you're talking about. But yeah I'd be happy to do test something out for you. I'm a yeast factory in my den right now.
 
bruin_ale said:
Something along these lines?
http://www.brewcrazy.com/hull-olive-oil-thesis.pdf
Click to expand...

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?
 
It did both, yeast viability and ester production. But yeah, it didn't really discuss growth - they were adding the olive oil during "storage".
 
passedpawn said:
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?
Click to expand...

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 :ban:) 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.
 
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. :eek:

Smuttynose_Farmhouse_Saison_yeast_w_violet_stain.jpg
 
passedpawn said:
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.
Click to expand...

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

Altbier_Yeast_Fuchschen_.jpg


Altbier_Yeast_Fuchschen_200x.jpg
 

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