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Old 11-15-2012, 02:10 PM   #1
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Default Help selecting a hemocytometer and microscope

For counting yeast cell density and viability what would be the most appropriate tools?

With a few searches on this site it seems that the $20 Hemocytometer, and an inexpensive microscope with 400x magnification are adequate, but I would like to hear about peoples experience with cell counting. What tools and techniques work, and which ones just don't cut the mustard.

Hemocytometers seem to range from $20 to over $200! For homebrewing I would imagine counts that are within a factor of two would be adequate. Would there be any need in a brew lab for the expensive hemocytometer?

The two microscope I have my eye on now are both from Amscope. One is about $80, monocular, fixed stage, up to 1000x, The second is $165, binocular, mechanical stage, up to 2000x.

$80 microscope
$165 microscope

$20 Hemacytometer
$200 hemocytometer

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Old 11-16-2012, 12:58 PM   #2
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I have that binocular Amscope, it works great for cell counting and is a very reasonable price. Mine goes up to 1000X, I can't imagine trying to use it at 2000X. Be very careful when installing the eye pieces that you don't get any lint or dust in. It is impossible to get out once any gets in, at least in my experience. My hemocytometer was $30 shipped I think, works fine.
There are some pics in my gallery taken from an iphone at one of the eye pieces. What you see in the scope is much clearer than what the pics show but it gives you an idea.

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Old 11-16-2012, 01:23 PM   #3
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That's good to know. It's nice to hear from someone that has real experience with very similar, if not the same, items. Thanks!

A picture speaks 1000 words. I know what you mean about the image not being quite representative with a cell phone. I took some pictures through my brother's Olympus scope and it just doesn't do it justice. It looks like the yeast pictures were taken at about 400x (10x eyepiece with a 40x objective) is that right?

One of the reasons that I am getting a microscope is to control my pitch rate from slurries. I noticed some other pictures in your gallery labeled "Re-used Slurry Viability" "Commercial Yeast Viability" and "Inoculation Rate Vs. Doublings"

Where did you get those graphs?

My experience is very limited, but here is what I have found so far:
http://woodlandbrew.blogspot.com/201...ing-cells.html

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Old 11-16-2012, 03:37 PM   #4
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Yes, they were taken at 400X. 400X shows a grid full view that you would count. 100X shows full view all 25 grids to see if you have even distribution.

I made those graphs with data from Jamil's calculator and info from the Yeast book.

More often than not, when I make a stir-plate starter using Jamil's calculator, I get ~75% of the cells the calculator predicts. So, I've been making bigger starters than it recommends, then do a cell count prior to pitching and use only as many cells (by volume) as the pitching rate I'm using dictates.

The hardest part about cell counting is figuring out what dilution to use and then measuring it accurately. I use 2 mL and 10 mL pipettes for starters and a 100 mL burette for slurries. The 1 mL pipette I got for taking a 1 mL yeast sample couldn't suck up starter slurry, the opening in the tip is too small so I have to use the 2 mL.

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Old 11-16-2012, 03:45 PM   #5
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I've been doing this for a while. Here's a link to a thread chock full of useful information. Good luck with your endeavors.

http://www.homebrewtalk.com/f163/pic...-scope-273342/

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Old 11-16-2012, 05:11 PM   #6
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dstar,
That's some good information. I have had a hunch that the calculators may have some inaccuracy, but wasn't sure how much. The 75% discrepancy you see is good to know. Have you seen a deviation between yeast strains and the accuracy of the calculated starters?

passedpawn,
You started one epic thread! I spent about 30 minutes reading it, but didn't get to the end. It is actually one of the sources I used to come to the equipment that I am considering.

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Old 11-16-2012, 06:20 PM   #7
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Quote:
Originally Posted by WoodlandBrew View Post
Have you seen a deviation between yeast strains and the accuracy of the calculated starters?
I've observed WLP001 multiple times and that is where I can confidently say 75%. I've used the WLP029, WLP380, WLP838 and Wyeast 1007 strains once each and seen ~75%

Under the same conditions, Wyeast 1098 and Wyeast 1728 grew to what Jamil expected. I thought starter size may have something to do with it since the 1098 was the first time I've seen Jamil numbers and it was a 2L starter (all others were 5L). Alas, the 1728 was also a 5L so the starter size theory can't be confirmed.

I've been playing with the Brett Trois strain from White Labs (WLP644) lately and it grows like crazy. 2.5 times the doublings of what Jamil predicts, after repeated starters.
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Old 11-16-2012, 09:37 PM   #8
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$165 for a binocular microscope that does 2000X sounds too good to be true. The 2000X is what we call 'empty magnification'. IOW instead of having a small blurry yeast cell you have a big blurry yeast cell. 1000X is more than you can get in an optical scope without oil immersion (is the 100X objective oil?). Empty magnification has sold lots of microscopes and telescopes to the unsuspecting.

Having said that you don't need anything like 1000x to count yeast cells. 100X is fine. At the price, if the thing performs decently at 100X it may be a good buy.

I usually advise people to try to find a good used microscope such as the ones used by medical students. These are somewhat over $1000 new and you ought to be able to find one in good shape for a couple of hundred. If you want to impress your friends with empty magnifications you can always buy a 20X eyepiece.

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Old 11-16-2012, 09:43 PM   #9
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Thanks AJ,

That's what I was afraid of. The 2000x did sound to good to be true. The 100x is an oil immersion objective. The most attractive things with the more expensive scope are that it is binocular and has a mechanical table.

The reviews are good on amazon. I'm hoping the 400x is good enough for counting cells and checking viability with methylene blue or trypan blue. If the higher powers aren't to blurry maybe I'll be able to see bacteria, however I hear plating is a better test for bacteria.

The 2000x is actually from a 20x eye piece and the 100x objective... maybe I need a 25x eyepiece to really impress my friends! ha ha.

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Old 11-16-2012, 10:05 PM   #10
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Yes, I saw those Amazon reviews. I'm still scratching my head. An oil immersion capability in a binocular scope for $165 is really unbelievable. You'll see what you'll see when you get it but you should be able to use it for counting in any case.

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