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Old 12-04-2012, 03:12 PM   #11
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Why count through inference when you can actually count?
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Old 12-04-2012, 03:27 PM   #12
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Speed.
Have you tried imageJ for automated counting (with scope/camera)? I've been meaning to give it a try.
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Old 12-04-2012, 04:40 PM   #13
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No, I haven't but I believe Kai has.

 
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Old 12-10-2012, 03:39 PM   #14
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I haven't used ImageJ for automated counting yet. But I use it for manual counting.

As for a practical alternative to counting cells, I would say pitching yeast by weight. Unfortunately there is some strain and obviously slurry dependency. But for well flocculating strains, where you get stable sediment, I found the cell count to be 3-4 Billion cells per gram in clean sediment.

I actually suggest that even brewers who have a microscope may want to establish a correlation between slurry weight and cell count since counting cells takes time and you may want to skip it at some point.

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Old 12-19-2012, 04:01 PM   #15
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Image J for manual counts is very nice. Especially for documenting results and checking your work, but for most cell counts I just use a clicker. The automated cell counting looks like it would take quite a bit of tweaking to get consistent results. The automated bit might be useful if you had weeks worth of counts to perform. Maybe months.

Some of my slurries are 1 billion cells per gram, others are 250 million cells per gram. A white labs vial I counted recently was 2 billion.

I use a microscope and hemocytometer. It works great and has opened my eyes to a whole new part of the brewing process. The variation from one slurry to the next, but my day to day results are very consistent, so I feel I am getting accurate results. but of course, the proof is in the pudding, or in this case I suppose, beer.

Here is the microscope and some pictures of yeast:

http://woodlandbrew.blogspot.com/201...icroscope.html
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Old 12-19-2012, 04:09 PM   #16
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If time isn't an important factor, you could count CFUs using agar plates. But personally as much as I hate counting cells with a hemocytometer, I hate making serial dilutions, plating, and counting colonies even more.

There are good ImageJ macros out there for semi-automated cell counting. It works well as long as you don't have a lot of yeast-sized debris in your sample.

 
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Old 12-19-2012, 04:09 PM   #17
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Quote:
Originally Posted by passedpawn View Post
Why count through inference when you can actually count? I've seen these Amscope microscopes for sale cheap (~$200 or less?). Cell morphology isn't possible but simple counting certainly is. A capable hemocytometer can be bought new on Ebay for $30.
I agree. That's the setup I have and it's worked well for me. It's fairly fast too. I can get down to about 5% standard deviation in about 15 minutes of work including dilutions and staining.

Here is a side by side comparison of the $30 to a $260 hemocytometer:
http://woodlandbrew.blogspot.com/201...e-by-side.html
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Old 12-19-2012, 04:11 PM   #18
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Quote:
Originally Posted by ipsiad View Post
If time isn't an important factor, you could count CFUs using agar plates. But personally as much as I hate counting cells with a hemocytometer, I hate making serial dilutions, plating, and counting colonies even more.

There are good ImageJ macros out there for semi-automated cell counting. It works well as long as you don't have a lot of yeast-sized debris in your sample.
Agreed and Agreed.

Is counting CFU's more accurate than a hemocytometer? I've considered it, but wasn't sure if it would really be worth it. With the Hemocytometer you are diluting 20:1 or 40:1 but with plating is serial diluting to something on the order of 1E6:1
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Old 12-19-2012, 04:25 PM   #19
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Agreed and Agreed.

Is counting CFU's more accurate than a hemocytometer? I've considered it, but wasn't sure if it would really be worth it. With the Hemocytometer you are diluting 20:1 or 40:1 but with plating is serial diluting to something on the order of 1E6:1
I'm sure some microbiologists out there would disagree with me, but I think the hemocytometer plus something like methylene blue is as good or better than counting colonies for exactly the reason you brought up. There's some error rate with methylene blue, but I think it's lower than the error you introduce by diluting something 1E6:1 or more.

In order to get good numbers from counting colonies, your starting sample and serial dilutions have to be perfect. And even when you do it perfectly with lots of vortexing, perfectly calibrated pipettes, etc. there's still variability so people do everything in triplicate and average the results. Not worth it for these purposes, IMO.

 
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Old 12-19-2012, 06:05 PM   #20
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Originally Posted by spenghali View Post
Is there a way to get an accurate cell count for pitching rates without the use of a microscope? I am looking for something other than a pitching rate calculator such as Mr. Malty.
Why? If you're using fresh yeast and pitching the amount that Mr. Malty recommends, you really can't go wrong. I haven't heard anyone yet complain that it didn't produce good results. You could always pitch a little more than what Mr. Malty recommends if it's a concern. Do you want a microscope just to see how many are actually there?

I'm just curious as to why you're looking for something other than a pitching rate calculator.


Edit: After reading this, the tone didn't come through - I really am just curious. No disrespect intended....
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