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Old 12-13-2012, 02:28 PM   #11
jerrodm
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Originally Posted by WoodlandBrew View Post
Has any one done cell counts to verify this? It's a common statement I hear, but my experience is that there is an even mix of live and dead cells between the layers. At one point Wyeast promoted this technique, but it seems they have removed it from their site.
I certainly haven't, and can't claim to have any specific knowledge about the ratio of living to dead yeast in different layers. I looked at your work, and it's convincing enough to me. I was speaking from the perspective of cleaning the yeast of hop material, hot and cold break, etc--the other things that get deposited in the bottom of your carboy. I frankly don't worry too much about the ratio of living to dead yeast--don't yeast often eat their dead buddies anyway, one reason that people use dead yeast as nutrients? As long as there's a sufficient amount of live cells (something your starter should confirm), you should be good to go.

I hadn't thought about the bacteria issue...that's an interesting point, and one I suppose you would want to consider when pitching. I've never gotten an infection from repitching washed yeast (although that doesn't mean that I couldn't!), so I've never considered it an issue. If higher bacteria concentrations are present in the top layer, would that argue for pouring off the top layer and pitching the bottom layer instead? Interesting thought.
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Old 12-13-2012, 02:28 PM   #12
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Originally Posted by pabloj13 View Post
It totally depends on the yeast. The sticky gives fantastic directions for washing and getting really nice cell counts (verified by microscope). However, confusion sets in in cases (like this one) where the yeast flocs out really quickly. The creamy white layer is the best layer for live cell counts. Most of the time that is the stuff that stays in suspension, but not always.
I'm glad someone has done cell counts and got good results. That's comforting. However, I've done cell counts on four different strains, and not seen the variation. They were: WLP004, WLP566, S-04, and EC-1118.

Here are the details on the first three:
http://woodlandbrew.blogspot.com/201...g-exposed.html

I don't remember seeing the cell counts in the yeast washing sticky. Could you please point me toward that?
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Old 12-13-2012, 02:35 PM   #13
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Quote:
Originally Posted by WoodlandBrew View Post
I'm glad someone has done cell counts and got good results. That's comforting. However, I've done cell counts on four different strains, and not seen the variation. They were: WLP004, WLP566, S-04, and EC-1118.

Here are the details on the first three:
http://woodlandbrew.blogspot.com/201...g-exposed.html

I don't remember seeing the cell counts in the yeast washing sticky. Could you please point me toward that?
The cell counts weren't in the sticky. They were in my hands. I didn't save the results, but I can do it again next time I wash.
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Old 12-13-2012, 02:37 PM   #14
pabloj13
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Quote:
Originally Posted by WoodlandBrew View Post
I'm glad someone has done cell counts and got good results. That's comforting. However, I've done cell counts on four different strains, and not seen the variation. They were: WLP004, WLP566, S-04, and EC-1118.

Here are the details on the first three:
http://woodlandbrew.blogspot.com/201...g-exposed.html

I don't remember seeing the cell counts in the yeast washing sticky. Could you please point me toward that?
I just read that post. Can you describe when you took the samples? (i.e. did you take cell counts from the stuff left behind in the fermenter?). It's interesting.
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Old 12-13-2012, 02:39 PM   #15
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Quote:
Originally Posted by jerrodm View Post
I was speaking from the perspective of cleaning the yeast of hop material, hot and cold break, etc--the other things that get deposited in the bottom of your carboy. I frankly don't worry too much about the ratio of living to dead yeast--don't yeast often eat their dead buddies anyway, one reason that people use dead yeast as nutrients? .
Yes, dead yeast makes good yeast nutrients. Also you are right that the viable cell count is more important than the viability. They are often used interchangably which can lead to confusion. Where it can be a problem is when the yeast is rinced with water the final volume is much smaller than the original. So if the viability is low, there are very few live cells.
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Originally Posted by jerrodm View Post
As long as there's a sufficient amount of live cells (something your starter should confirm), you should be good to go..
This seems true for the most part. One of my recent slurries was 1% when I harvested it. Out of curiosity I made a starter out of it to see if it would take off or if the bacteria would dominate. Because I used the actual live cell count to determine the pitch rate, and the yeast had just been harvested it took right off. I suppose if I had assumed 90% viability I would have seen very little activity.
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Originally Posted by jerrodm View Post
If higher bacteria concentrations are present in the top layer, would that argue for pouring off the top layer and pitching the bottom layer instead? Interesting thought.
After crashing the slurry I pour off the top and fill it back up with clean water. That lowers the alcohol level which is part of what protects the yeast, so I'm not really sure if this is the best technique, but time will tell.
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Old 12-13-2012, 02:42 PM   #16
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I just read that post. Can you describe when you took the samples? (i.e. did you take cell counts from the stuff left behind in the fermenter?). It's interesting.
Those experiments were based on slurries that had been in the refrigerator for a period of time and then crashed in test tubes. A pipette was used to remove samples of each layer without disturbing them. I'm collecting data now on an active fermentation and am seeing the same mix of live and dead cells in the suspended yeast.
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Old 12-13-2012, 02:44 PM   #17
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The cell counts weren't in the sticky. They were in my hands. I didn't save the results, but I can do it again next time I wash.
Awesome! I wonder why we are seeing different results. It would be interesting to see what the difference was in viability.
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Old 12-13-2012, 02:50 PM   #18
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Awesome! I wonder why we are seeing different results. It would be interesting to see what the difference was in viability.
For sure. For me the biggest reason I wash is just to get rid of most of the hop debris and trub since I just put EVERYTHING into the fermenter. There is no question, though, that people have success direct pitching slurry. I also take my crashed yeast and freeze them at -80 with 15% glycerol so I can keep them long term, so I want them as clean as possible. I'll be washing again soon. Now I have an experiment to do. I love experiments.
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Old 12-13-2012, 03:09 PM   #19
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I love experiments.
Me too! The sciency bits (that's a technical term) are the parts that I enjoy the most. I have one fermentation finishing up soon, and another in about a week. I'll re-read the sticky and follow it the best I can. It would be wonderful to be able to actually wash out 90% of the dead cells. It would make determining the number of viable cells much more accurate without a microscope. (Viability and thick cell density [likely driven by protein content] are the two biggest factors working against those not using microscopes, but that is a whole different discussion)
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Old 12-13-2012, 05:40 PM   #20
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good info. Im still left wondering how to proceed....

Should I mix the slurry up again and wait for it to resettle to gauge the results before rewashing?

Or should I simply dump the top two layers of dark murkiness and clear beer and retain the bottom creamy white layers?

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