theveganbrewer
Well-Known Member
Thanks. 16oz of glycerine or the big gallon jug? I'm having a hard time understanding the scope of this process.
Do you know how to make a yeast starter? Then why not farm yeast and freeze it?
- Thread starter BBL_Brewer
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BBL_Brewer
Well-Known Member
It's up to you, but I like the 4 oz canning jars. They're easier to store IMO and you can get them almost anywhere.
Golddiggie
Well-Known Member
Unless you plan on freezing a LOT of yeast, I'd get the quart bottle first. You'll be diluting it, so that will go a long way. You can plan the next purchase according to how fast you use the first one up.
Yes, 50 ml centrifuge tubes from Amazon are ideal. Here is the link for smaller quantities:
http://www.amazon.com/dp/B0018MQ9TK/?tag=skimlinks_replacement-20
I'm going to post my article soon. It has sources for glycerin and plastic tubes. Also a detailed discussion of the whole process summarized from this thread.
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Golddiggie
Well-Known Member
I have over a dozen of these: http://www.amazon.com/gp/product/B005CBFATS/?tag=skimlinks_replacement-20
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theveganbrewer
Well-Known Member
You guys are awesome. It's like a live chat with the yeast customer service department.
I just submitted the article on Freezing Yeast. I don't know if its visible to just me or everyone but it is in the Technical Article section right now. I don't know what kind of file the moderator wants but I did it as a PDF file. Hopefully you all like it and agree with my distillation of the methods. In any case, what I proposed should work great and many successful modifications on the theme are possible.
theveganbrewer
Well-Known Member
So thanks to your links I found out those white lab tubes are called baby soda bottles, which opened up a ton of links to buy them.
Golddiggie
Well-Known Member
They're smaller than the ones I linked to. I'd rather have more slurry in the tube than less...
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BBL_Brewer
Well-Known Member
If you submitted it for a membership we can't see it, it's hidden.
Do you know if its ok to upload it here as well, or does that make it a moot point for him to post it in the technical articles section?
theveganbrewer
Well-Known Member
It'd be nice to have it here too 
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BBL_Brewer
Well-Known Member
Don't know on both accounts.
coryforsenate
Well-Known Member
I regularly have to make freezer cultures of bacteria in my lab. I haven't perused the entire thread to see if this has been noted, but you'll get marked viability increases the faster you freeze cells because of both the ice crystals being smaller and the presence of a cryoprotectant (glycerol) preventing the formation of the standard hexagonal crystal lattice.
We use 50% v/v glycerol and liquid nitrogen. Liquid nitrogen is a bit hard to come by for most folks, but dry ice will work to achieve faster freezing.
Also, at least in bacteria, thawing quickly reduces cell viability. When we remove cultures from our -80C freezer, we put them on ice until they thaw fully.
We use 50% v/v glycerol and liquid nitrogen. Liquid nitrogen is a bit hard to come by for most folks, but dry ice will work to achieve faster freezing.
Also, at least in bacteria, thawing quickly reduces cell viability. When we remove cultures from our -80C freezer, we put them on ice until they thaw fully.
orangehero
Well-Known Member
Works better opposite for yeast: slow cooling, rapid thaw. Yeast will continue to build up trehalose which also serves cryoprotectant functions.
coryforsenate
Well-Known Member
Interesting. Just another way yeast and bacteria are different.
Brewitt said:
If you would like too, I can put your write up in the wiki. I have done a few wiki articles myself. Let me know and send me a word doc link pm if you do want it there. This is a wealth of info and should be there where we can link it to farming and washing threads as well. If I do this, you lab guys will need to take some more photos to make the article more like a magazine. I really need to update my pressurized fermentation wiki article as well. Anyways, it would be fun and beneficial to have it done in wiki. Great thread guys.
WortMonger said:
Thanks. I'm not familiar with the wiki. I submitted it to the technical thread for approval. The directions say that it needs to not be submitted anywhere else. I don't know if that also applies to anywhere on this forum. I also don't know how long it takes to get approved.
Forkhead
Active Member
What are people mixing the glycerol with to freeze? I used fresh wort because I was worried about osmotic stress, but it was a pain because the yeast was making CO2 when they warmed up and made things very messy. I actually had a yeast bomb go off in my apartment!!! What a disaster!
Will putting yeast in straight water be bad for viability? I know with mammalian cells they'll all die instantly in water.
Will putting yeast in straight water be bad for viability? I know with mammalian cells they'll all die instantly in water.
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BBL_Brewer
Well-Known Member
I decant off most of the spent wort after the starter is done and then add sterile glycerol/water solution. So it's mostly water, glycerol and yeast with a little bit of spent wort. I was wondering how the wort thing would work out for you. Even with my method I still have to do some venting when I pitch a jar. When I swirl to get the yeast in suspension I get a little off gassing.
I agree with BBL_Brewer. I have typically made a dense slurry leaving in whatever spent wort is necessary to have a slurry instead of packed cells. I then bring up in an equal volume of glycerol prepared in water. The spent wort (if fully fermented out) should not cause a problem. Washing yeast with water is not particularly detrimental to viability, so the water in the glycerol should not be a big problem. Between the cell wall and effective ion pumps, they do a respectable job of maintaining their osmolarity.
Forkhead
Active Member
OK, thanks for the info! I'm growing up another culture to repeat the freeze/thaw. I'd like to optimize the procedure a little bit. I'll try water+glycerol instead of wort+glycerol. I'm also going to test a few other parameters.
It looks to me that these are thin walled polycarbonate tubes. That is not really a problem but they certainly won't have the durability of White Labs tubes which are very thick walled. They are about the same diameter and hold only 40 mls. They may have problems during freezing and thawing but that remains to be seen. I believe you would be better off with the centrifuge tubes which are quite durable and close tightly.
Does anyone know differently?
Golddiggie
Well-Known Member
The tubes I linked to are THICK walled (along the lines of those from White Labs)... These also close VERY tightly. I have a couple of packages of them on hand, so it was easy to check.
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rockinmarty
Big Member
@Golddiggie just found the same size cheaper if anyone is interested here is the link
http://www.stevespanglerscience.com/product/test-tubes/134
I love this thread, but that gives me one more project to work on that I don't have the time for
http://www.stevespanglerscience.com/product/test-tubes/134
I love this thread, but that gives me one more project to work on that I don't have the time for
The tubes I linked to are THICK walled (along the lines of those from White Labs)... These also close VERY tightly. I have a couple of packages of them on hand, so it was easy to check.
Point taken. I apologize for misleading and misinterpreting. I saw people saying in reviews that these were like miniature soda bottles. Sounds like I was wrong and that these are like the the clear 40 ml polycarbonate tubes we use in laboratories. Those should be perfect.
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Brewitt said:
They ARE miniature soda bottles, though possibly not in the sense you're thinking. They are called preforms, and are typically used by the bottling facility by expanding them to full size by heating them and blowing air into them (an effect that resembles glass blowing). They are exactly what White Labs uses, and ARE fairly thick-walled because they contain enough material to make a typical 2L bottle... the walls get thinner as air is blown into them to "stretch" to full-size.
They are popular because they are already manufactured on a massive scale for the beverage industry, making them a very cheap (literally pennies when bought in bulk) alternative to purpose-made labware. Notice how the caps are *identical* to soda bottles? It's not a coincidence.
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