Home Brew Forums > Home Brewing Beer > Fermentation & Yeast > Do you know how to make a yeast starter? Then why not farm yeast and freeze it?
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Old 09-28-2012, 06:00 PM   #181
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I use trypan blue to measure viability, but I don't know where you can buy it other than scientific supply stores like Sigma-Aldrich. I don't know if they'll sell to an individual user not affiliated with an organization. I think you can also use methylene blue, which you may have an easier time finding.

If you want to do it, you take a small sample of your yeast and dilute it in trypan (methylene) blue and place on the microscope. Live cells exclude trypan (methylene), dead cells will look dark blue.


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Old 09-28-2012, 08:50 PM   #182
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I use trypan blue to measure viability, but I don't know where you can buy it other than scientific supply stores like Sigma-Aldrich. I don't know if they'll sell to an individual user not affiliated with an organization. I think you can also use methylene blue, which you may have an easier time finding.

If you want to do it, you take a small sample of your yeast and dilute it in trypan (methylene) blue and place on the microscope. Live cells exclude trypan (methylene), dead cells will look dark blue.
Cool. That sounds easier than the way we did it in micro back in the day. Thanks.


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Old 09-29-2012, 12:45 AM   #183
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Colony formation is a lot more accurate.
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Old 09-29-2012, 01:47 AM   #184
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Colony formation is a lot more accurate.
I agree colony forming units (CFU) is the most accurate way to measure viability. The problem is that it takes 24-48 hrs for colonies to grow up. Meanwhile, what do you do with your starter? It'd be nice to have a real time measurement of viability. As far as I can find, no one has directly compared viability measurements by CFU or trypan/methylene blue to see if they have similar accuracy.

If I have time, I'll pour some plates and grow up colonies to compare CFU with my trypan blue measurements. My yeast got delivered today, I'll have some results in just over a week.
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Old 09-29-2012, 04:13 AM   #185
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I really don't want to get into plating. Although it would save some money to streak once in a while. Even if staining is less accurate, it's still better than flying bilnd and a lot less hassle.
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Old 10-03-2012, 03:48 PM   #186
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Well, the experiment has begun. I made my starter last night. I decided to use California Ale yeast from White Labs (WLP001). Before pitching the yeast, I took out a small aliquot to count on a microscope and hemocytomter, and measured viability by Trypan Blue exclusion. The total # of viable yeast cells in the vial from White Labs was ~75 billion and this correlated to ~85% viability.

I was expecting the total # to be closer to 100 billion, but I guess White Labs doesn't actually put that many in each vial. I also plated the cells on YPD plates to compare the numbers from the hemocytometer/Trypan with actual colonies. It'll take ~24 hrs for the colonies to grow up. I'll post again when I get those results. Meanwhile, I'll keep growing up more yeast in the starter to test various freezing conditions.
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Old 10-03-2012, 04:06 PM   #187
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Well, the experiment has begun. I made my starter last night. I decided to use California Ale yeast from White Labs (WLP001). Before pitching the yeast, I took out a small aliquot to count on a microscope and hemocytomter, and measured viability by Trypan Blue exclusion. The total # of viable yeast cells in the vial from White Labs was ~75 billion and this correlated to ~85% viability.

I was expecting the total # to be closer to 100 billion, but I guess White Labs doesn't actually put that many in each vial. I also plated the cells on YPD plates to compare the numbers from the hemocytometer/Trypan with actual colonies. It'll take ~24 hrs for the colonies to grow up. I'll post again when I get those results. Meanwhile, I'll keep growing up more yeast in the starter to test various freezing conditions.
Did you use the $24 amazon scope to view the cells with the hemo?
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Old 10-03-2012, 04:40 PM   #188
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I first counted at home using the $24 microscope from Amazon, then I took the cells into work to count using a real microscope. I was able to get pretty close using the $24 scope, but it was pretty difficult, and I have many years of experience counting cells on microscopes. There's no way you could do a viability measurement on the $24 scope because it was impossible to distinguish live from dead cells.

Another tip for anyone wanting to repeat this. I pitched a 35ml vial of White Lab yeast into 1.4L of wort, which is a 40-fold dilution. I had to dilute the aliquot that I took out of the starter another 20-fold before they were dilute enough to count on the microscope, which is a 800-fold total dillution. For plating the cells, I diluted the aliquot taken from the starter 100,000-fold so that hopefully there will be ~100 colonies per plate.
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Old 10-03-2012, 05:09 PM   #189
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Well, so much for a cheap scope. Thanks for the info.
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Old 10-03-2012, 05:13 PM   #190
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One concern less. Thanks for info.


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