Pics of Yeast under my new microscope

Williams 728
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kcpup said:
PassedPawn, would this microscope work ok?

Costco - Celestron Touch Screen LCD Deluxe Digital Microscope

Thanks for your input
Click to expand...

That would be great if it did, wouldn't it!

400x is perfect for counting in a counting chamber, so the magnification is plenty. All the nice features there make it look like the greatest deal ever seen in microscopy. Unfortunately, you know what they say about deals "too good to be true".

Those who seem to know about these kinds of things say that scopes like these are a waste of money because the optics are crap and only result in frustration. But if I were you I'd try get a live demo on one of those things, dial it to 400x (40x objective), and ask yourself if you could clearly resolve yeast cells on that screen. The screen resolution is going to GREATLY affect the usefulness of that scope.

They list a mechanical stage, which presumably allows you controls in X and Y, but at magnfication it might not give smooth movement and fine-enough adjustment to be useful. This would another thing to check.

I'm sceptical, but for $200 I'd absolutely buy one if it even barely worked. Besides, with the digital photo/video, it would also save you from the kludge I've been using to get pics of the cells.
 
passedpawn, if you're taking requests for your "to do" list, could you count the number of cells in a bottle conditioned beer - say a Bell's Amber or Pale Ale?
 
BTW, PassedPawn, if I buy a traditional microsope (as opposed to that LCD one I just asked you about) I was considering this as an add on.

Amazon.com: Celestron Digital Microscope Imager: Industrial & Scientific

This may not be high enough MP for you, but thought I'd pass it along. At the lower magnification (40x included), people are saying it does a decent job...
 
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jmf143 said:
passedpawn, if you're taking requests for your "to do" list, could you count the number of cells in a bottle conditioned beer - say a Bell's Amber or Pale Ale?
Click to expand...

All right. I'll be picking up a bells 2-hearted this wkend (just made a clone and want to do a comparison). I'll give it a good shake to get the yeast into suspension and do that. It only takes a minute (after I get the dilution right, but I suspect I won't have to do that with bottled beer). I'll probably mess around with video again and post some more of that.
 
passedpawn said:
All right. I'll be picking up a bells 2-hearted this wkend (just made a clone and want to do a comparison). I'll give it a good shake to get the yeast into suspension and do that. It only takes a minute (after I get the dilution right, but I suspect I won't have to do that with bottled beer). I'll probably mess around with video again and post some more of that.
Click to expand...

I should have been more specific - how about just from the dregs of the bottle, as that is what I'd be trying to culture.
 
jmf143 said:
I should have been more specific - how about just from the dregs of the bottle, as that is what I'd be trying to culture.
Click to expand...

I haven't gotten around to getting stains, so I can't determine viability. That would be a significant concern. But thanks for the reminder. Stains are dirt cheap online, and I think I'll get them for some fun during Christmas holidays.
 
I'm soooo jealous! I want a microscope! My wife thinks I'm even more of a nerd for asking for one for Christmas.
 
jmf143 said:
passedpawn, if you're taking requests for your "to do" list, could you count the number of cells in a bottle conditioned beer - say a Bell's Amber or Pale Ale?
Click to expand...

OK, saved my dregs from a bottle of 2-hearted this wkend.

Amount of dregs: 24ml (355ml in a bottle)
Cells per ml: 850,000
Total cells in dregs / bottle: 302 million
​
That sounds about right, too.
 
passedpawn said:
OK, saved my dregs from a bottle of 2-hearted this wkend.

Amount of dregs: 24ml (355ml in a bottle)
Cells per ml: 850,000
Total cells in dregs / bottle: 302 million
​
That sounds about right, too.
Click to expand...

Thanks for doing this passedpawn. I appreciate the work you did.

Is 302 million the total number of cells and not just the viable ones? You talked about getting stain so you could differentiate the cells - I'm assuming you haven't gotten it yet.
 
What type of stain are you looking to get? I count yeast a lot for viability and I am in the process of switching from methylene blue to alkaline methylene violet. The dye used to make the latter is methylene violet 3RAX. I have only really found it a Sigma Aldrich and they require you be a company or something order. I also believe that the dye is mixed into a glycine buffered alkaline solution. I made a little order mix up last time I sent something to the purchasing department, and neglected to include the dry dye in my order list. The cool thing is with the 1gram of dry dye you get a TON of solution to use. You simply take .1gram into 1L and 100ml of that stock is used to make 1L of actual die. So as you can see for $25 1gram of dye goes a real long goddamn way when you're using 1ml per sample. Your other option would be to order 100ml worth I think from White Labs. It's expensive compared to buying the ingredients in terms of what you get but it'd sure be easier.

My school has a very high end scope that I just found out has a sweet ass camera attached. As soon as this semester is over (its finals week now) my prof will show me how to not screw it up since it's like $50k, and then they'll set me free to take pictures of whatever I want.

If you need any help or info let me know.
 
jmf143 said:
Thanks for doing this passedpawn. I appreciate the work you did.

Is 302 million the total number of cells and not just the viable ones? You talked about getting stain so you could differentiate the cells - I'm assuming you haven't gotten it yet.
Click to expand...

Don't have dye yet. That was the total. Sorry, should have made that clear.
 
smokinghole said:
What type of stain are you looking to get? I count yeast a lot for viability and I am in the process of switching from methylene blue to alkaline methylene violet. The dye used to make the latter is methylene violet 3RAX. I have only really found it a Sigma Aldrich and they require you be a company or something order. I also believe that the dye is mixed into a glycine buffered alkaline solution. I made a little order mix up last time I sent something to the purchasing department, and neglected to include the dry dye in my order list. The cool thing is with the 1gram of dry dye you get a TON of solution to use. You simply take .1gram into 1L and 100ml of that stock is used to make 1L of actual die. So as you can see for $25 1gram of dye goes a real long goddamn way when you're using 1ml per sample. Your other option would be to order 100ml worth I think from White Labs. It's expensive compared to buying the ingredients in terms of what you get but it'd sure be easier.

My school has a very high end scope that I just found out has a sweet ass camera attached. As soon as this semester is over (its finals week now) my prof will show me how to not screw it up since it's like $50k, and then they'll set me free to take pictures of whatever I want.

If you need any help or info let me know.
Click to expand...

I was planning on buying methylene blue. Why are you considering going to the violet stain?
 
passedpawn said:
I was planning on buying methylene blue. Why are.you considering going to the violet stain.
Click to expand...

The reason for the switch was everything I keep reading about the accuracy of Methylene blue.

Rather than repeat the discussion in the easiest to reference source here's the link. Starting around page 142 is what I'm talking about. You may find other sections of the book interesting as well. Essentially it comes down to accuracy and ease of use. If you look at some of the data in the book you can see heat killed yeast still showed unrealistically high viability with methylene blue.

Brewing Yeast Fermentation Performance
 
So I stopped by my moms to find my dads old scopes. One is a reichert jung series 40 with a 100x and 200x objectives. The second one is a bausch & laumb khs (I think) with 40x,100x, 400x, and 1000x.

ForumRunner_20111213_193656.jpg
 
I just bought a microscope for at work for doing algae counts. I may pm some of you guys when I get around to playing with it and have questions.
 
So I was playing around with the scope. I thought it was messed up because I was seeing double. My wife said she isn't seeing double. Would lasik make this happen?
 
I just wanted to know if the scope was messed up or my eyes.

I always do that when I get drunk too.
 
Try adjusting the width between the eye pieces while looking thru it. Should merge the 2 drunken images. :)
 
I tried that. It got too wide (pause for that's what she said comment)

my other scope doesn't do double vision but it is only a 200x.

I did receive my hemocytometer in the mail the other day too. I need to clean the scope, its been in the garage for at least 4 years.
 
smokinghole said:
What type of stain are you looking to get? I count yeast a lot for viability and I am in the process of switching from methylene blue to alkaline methylene violet. The dye used to make the latter is methylene violet 3RAX. I have only really found it a Sigma Aldrich and they require you be a company or something order. I also believe that the dye is mixed into a glycine buffered alkaline solution. I made a little order mix up last time I sent something to the purchasing department, and neglected to include the dry dye in my order list. The cool thing is with the 1gram of dry dye you get a TON of solution to use. You simply take .1gram into 1L and 100ml of that stock is used to make 1L of actual die. So as you can see for $25 1gram of dye goes a real long goddamn way when you're using 1ml per sample. Your other option would be to order 100ml worth I think from White Labs. It's expensive compared to buying the ingredients in terms of what you get but it'd sure be easier.

My school has a very high end scope that I just found out has a sweet ass camera attached. As soon as this semester is over (its finals week now) my prof will show me how to not screw it up since it's like $50k, and then they'll set me free to take pictures of whatever I want.

If you need any help or info let me know.
Click to expand...

I found the dry dye here. 1g makes 1000 ml


But because of the small amount I need, the White Labs solution is probably the way I'll go. $12 for 50ml.


I think that while I'm waiting for that to come in the mail, I'll pick up some Methylene Blue (aquarium store).
 
Right 1g makes 1L of stock. Each 10ml of stock makes 100ml of dye. Sorry each 1g makes 10L of dye not 1000L. The stuff you're buying at White Labs is the diluted stock. Not the .1g diluted solution.
 
smokinghole said:
Right 1g makes 1L of stock. Each 10ml of stock makes 100ml of dye. Sorry each 1g makes 10L of dye not 1000L. The stuff you're buying at White Labs is the diluted stock. Not the .1g diluted solution.
Click to expand...

Thanks. I was unsure of the second dilution. I'm not a chemist, and I have very limited ability to do anything related to chemistry :eek: As much as I would enjoy adding a bunch of new glassware to my inventory of toys, I'm gonna just buy the ready-to-go stain from White Labs. Thank you so much for helping me out here.
 
You need a dye like trypan blue to label dead cells. Invitrogen sells this aka gibco, for like 20$ for 100ml I believe

Edit- never mind this has been discussed
 
Yeastgroupwithscale2cropped.jpg


WLp001-1.jpg


I finally got on my school's scope with the camera. You can see in the one with the dye the budding yeast are more concerned about budding and will not expel/metabolize the dye. They are alive and well they're just not concerned about anything but budding at that point.
 
They should be great, that damn scope is like $30,000. I want to see if I can take some florescent pictures. I just need to get a florescent dye that will bind to certain proteins on the cell surface or interior. This scope has a florescence attachment that can take some really cool pictures.
 
smokinghole said:
They should be great, that damn scope is like $30,000. I want to see if I can take some florescent pictures. I just need to get a florescent dye that will bind to certain proteins on the cell surface or interior. This scope has a florescence attachment that can take some really cool pictures.
Click to expand...

can you do phase contrast?
 
robh245 said:
I am looking at microscopes for this and wanted to see if anyone thinks this type of microscope would be sufficient to see yeast buds or even do cell counts for viability. It is a USB 40x to 800x but not oil immersion. I'm curious as to what kidn of quality I could expect.

Amazon.com: 800x 2mp 8-led Usb Digital Microscope with Handsfree Stand (Support Windows 7(support 32 Bit Only!)/vista/xp/2000/mac Os X 10.5 or Above): Camera & Photo

Thanks for any insight!
Click to expand...

You don't need oil immersion. That is usually only necessary for objective magnifications of 100x or greater. With a 40x objective (and 10x eyepiece), you have 400x which is terrific for counting on a hemocytometer.

With an oil immersion objective, you might really get a good closup look at yeast budding, but you'd see it without also. Cell morphology would be easier with oil immersion also.

That scope isn't going to do it for you. While the magnification is there, you need the type that is lighted from beneath (lamp -> condenser). Might want to read the reviews of anything you buy first.
 
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I've always been curious, do yeast by chance have an endoplasmic reticulum, or a golgi apparatus?

Just thinking back to some of my favorite terms from my biology days...
 

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