Setting up a yeast lab?

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Storing yeast, regardless, requires a little effort (very little, imo) building up step cultures occasionally. It takes minutes with stored vials of wort, etc., ready to hand. Very little hands-on time, in reality. Ideally with at least some repitched generations, to appreciate the reward of pitching something that can’t be bought. The best pitchable yeast possible. There’s really no need for home brewers to make it much more complicated than it needs to be.
 
Check out Sui Genesis Brewing (link below). Brian is a microbiologist by day and has some useful info on how to bring some of these techniques to the home scale without breaking the bank. Wild yeast collection and isolation, cell count and viability, clean workspace, etc. Lots of other interesting stuff on his site and YouTube videos, too.

https://suigenerisbrewing.com/
 
Yeast brinks seem to be the best way to store yeast. They need to be 100% oxygen free to be most effective.
Yes, can’t beat it, but it’s a vessel for storing harvested fresh yeast slurry for days, on the verge of being repitched, while the yeast cells are still at the highest levels of viability and vitality. Here’s a simple scaled-down home brew version:
 

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I do overbuild starters and save them in 250 ml Erlenmeyer flasks. The different yeasts seem to have the same amount of sediment and since it's an overbuild the trub is minimal. So I deem them to have the same cell count,and make starters accordingly. Every procedure needs an SOP in my brewery.
 
So I deem them to have the same cell count,and make starters accordingly.
The potential problem here, with large volumes of stored slurry, as the slurry ages, is cell counts and assumptions ignore a crucial part of the puzzle, vitality, which declines too over time. This can result in under pitching even if enough cells have been pitched. A case where pitching fewer healthier cells ferments better than more unhealthy cells. By starting at very small volumes, like 10ml, you force things in your favour.

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Thanks for sharing. I think my approach for lager yeast will be to stick with my 50ml vials (25ml slurry + 25ml glycerin/wort) but just use more. Take out six vials, thaw them out and put into 2 parallel starter routines. I know this is not the same as starting with one 10ml vial and growing it up but it is for convenience which has to have some sacrifices. I see 50ml vials as better than 150ml vials for freezing. If it all does not work well, it is just yeast, my time and some learning. Thanks for your perspective.
 
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Thanks for sharing. I think my approach for lager yeast will be to stick with my 50ml vials (25ml slurry + 25ml glycerin/wort) but just use more. Take out six vials, thaw them out and put into 2 parallel starter routines. I know this is not the same as starting with one 10ml vial and growing it up but it is for convenience which has to have some sacrifices. I see 50ml vials as better than 150ml vials for freezing. If it all does not work well, it is just yeast, my time and some learning. Thanks for your perspective.
Bear in mind that freezing, regardless, is going to kill and knacker yeast cells at a high rate determined by a number of factors and that they won’t be conditioned well for fermenting wort. Adding more won’t change much and most likely promote an extended ‘lag phase’ with associated risks, especially for a lager, in terms of partial infection and yeast stress leaving subtle off flavours. Try to do a meaningful comparison with repitching freshly harvested yeast. I find a lager yeast cake kept cool under beer in primary is good for repitching directly for at least 2 weeks after fermentation finishes.
 
My problem is that I only brew lager a few times a year. And some ale styles months apart. So repitching is not a viable option for me. I see freezing the yeast as a 'long term repitch' in some way. It is my only way to have a chance at keeping yeast and using them further down the line. So far, the ales have been acting well. I have done a few lagers that have had long ferments, but I think there was a severe underpitch involved as I only started with two vials and fermented the beers at 48F with 2 bar of pressure. Pretty stupid looking back! Which is why I am thinking to start with six vials out of the freezer and see if my outcome is improved.

Pitfalls would be that I am moving around and pitching more dead yeast cells. But I also gain more live yeast cells. This is a frustrating part of the hobby for me as I do not have the time or inclination to properly prop up a yeast cake for lagers. It is still a hobby and I do not follow lab procedure like you do with a microscope etc... Just trying to find the best way forward that still has a percentage of success.

So given all of that, can you think of a better way for homebrewers to achieve long term repitching? Thanks.
 
So given all of that, can you think of a better way for homebrewers to achieve long term repitching?
It's impossible to maintain sufficient viability and vitality of wet yeast for direct pitching without feeding them regularly, which is a lot more hassle than stepping up from small volumes. Note if a procedure causes high rates of mortality in yeast cells, imagine what it does the the cells which survive. I'll only go to the effort when I plan to brew at least a few batches back-to-back, so I can repitch the yeast at their best. Binge brewing. But, as I've noted, if you already have the required sterile wort step ups frozen in bottles ready to go, it's just a little time management with minimal hands-on. I don't even bother with a stirplate anymore, just leave starters on the countertop in my brew room and swirl them whenever I pass.
 
I take the frozen 50ml vials out to thaw for 24 hours then put them in 200ml of 1.020 wort for a few days then add 2L of 1.035 wort and let that ferment out. Crash, decant then pitch. So not really direct pitching and hopefully a fair amount of new growth. For the lager example I would do two of these at the same time but starting with three vails each instead of one. Proportionality is off but hopefully more cells in the end.

Maybe I need to invent an autofeeder that sits in the fridge and keeps the yeast fed on a schedule? :)
 
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When I used to keep a sourdough starter and feed it daily, the thought of occasionally adding a tablespoon or two of LME to a pitchable amount of brewer's yeast slurry did cross my mind. It might at least keep them pitchable for longer than weeks.
 
When I used to keep a sourdough starter and feed it daily, the thought of occasionally adding a tablespoon or two of LME to a pitchable amount of brewer's yeast slurry did cross my mind. It might at least keep them pitchable for longer than weeks.
I feed my starter every week or 2 when it’s in the fridge
Sorry, sourdough.
 
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Has anyone actually measured cell count and viability in their frozen cultures? Pretty easy with a relatively small financial investment. I've never bothered because I get pretty good results from stepping up a single 50ml conical for a 2L starter (lagers, mostly).

To me, a home yeast lab means: microscope, hemocytometer, glycerine, and 50ml conicals. Not much else needed at this level (unless you plan to ranch wild yeasts).
 
Yes and, judging from the extended “lag phase”, vitality was pretty poor too. Presumably why commercial yeast suppliers don’t sell yeast in this frozen format. I suspect my lager fermentations are done by the time yours get started. My fermentations are predictable so I can plan ahead with confidence. I’m in control of the process with my method. Yours is more “wait and see” with fingers crossed. If you disagree, contact Chris White and tell him about your exciting discovery.
 
There are a lot of technical considerations when freezing and thawing yeast such as osmotic pressure of solution and speed of freezing and thawing. I have decent wet-lab education, training, and experience which carries over well to the home brewing. I haven't specifically measured my cell counts and viability, but get decent enough fermentation timecourses and flavor profiles in beer.

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So, it's likely not as simple as "freezing yeast doesn't work." There will be a heavy influence of the technical aspects of the lab procedures that one implements. I think one key aspect everyone could do is to freeze yeast slowly (about -1C/min) by placing in an insulated container in the freezer and thawing rapidly about about 30-35C.
 
In a previous life, I did QC/QA/product documentation for a company that sold frozen micro-organisms, and I can assure you that our methods were a lot closer to what McMullan describes than almost any of the "how to reuse your yeast" advice you will find on the internet. If you're happy with the results you get with one of the various shortcuts that homebrewers use, then by all means keep doing that. But if you're going to go to the trouble of setting up something worthy of being called a lab then you might as well do things right. And "right" means preserving a population of healthy cells and scaling up from a single one of those healthy cells for each use.
 
I don't know about yeast but in the food industry the faster(colder) the food is frozen the less ice crystals form. I installed doors in these 20 below 50 mph wind freezers.
 
It's pretty well established that rapid freezing promotes higher rates of ice crystal formation in living yeast cells. Regardless of food processing and rock formation. The scientific literature goes back decades with more reviews you could shake a stick at. Ice crystals are not good for living cells either. Once frozen, the yeast cells are under stresses associated with dehydration too. In fact, thawed frozen yeast are in worse shape than hydrated dry yeast. They are knackered and need to be carefully selected for post thawing. This is why my view, for home brewers generally, at least those who want to store liquid yeast strains, is shifting towards storing small aliquots of high-quality yeast in the fridge. I can't say it's a viable alternative for long-term storage over years, I'm genuinely skeptical about that. But I can say it's a better, easier option for most home brewers, at least up to about 18 months so far. I stand by my original reply to the OP, that freezing yeast cells is technically demanding involving a lot more than simply freezing tens of mls of slurry with glycerol then thawing to pitch or make a large starter. That's not sound microbiology or good brewing practice at all therefore not something I can endorse, because it's simply not something I'd do myself.
 
It's pretty well established that rapid freezing promotes higher rates of ice crystal formation in living yeast cells. Regardless of food processing and rock formation. The scientific literature goes back decades with more reviews you could shake a stick at. Ice crystals are not good for living cells either. Once frozen, the yeast cells are under stresses associated with dehydration too. In fact, thawed frozen yeast are in worse shape than hydrated dry yeast. They are knackered and need to be carefully selected for post thawing. This is why my view, for home brewers generally, at least those who want to store liquid yeast strains, is shifting towards storing small aliquots of high-quality yeast in the fridge. I can't say it's a viable alternative for long-term storage over years, I'm genuinely skeptical about that. But I can say it's a better, easier option for most home brewers, at least up to about 18 months so far. I stand by my original reply to the OP, that freezing yeast cells is technically demanding involving a lot more than simply freezing tens of mls of slurry with glycerol then thawing to pitch or make a large starter. That's not sound microbiology or good brewing practice at all therefore not something I can endorse, because it's simply not something I'd do myself.
I might have missed it McMullan, but are your aliquots simply good yeast under some amount of wort? Loose cap?
 
I might have missed it McMullan, but are your aliquots simply good yeast under some amount of wort? Loose cap?
These ones are straight from a White Labs’ WLP007 pack. ‘Outsourcing’ the lab work, making it incredibly simple. They know what they’re doing. Or make a nice clean starter with your strain(s) of choice and store the same way. The cryovial caps are screwed on tight. They can handle a little pressure, but I haven’t noticed any building up. I recommend just pitching one vial in a 50-100ml first step starter, for simplicity, but I actually just use a loop’s worth to inoculate 10ml sterile wort then step up to 100ml, 500ml, for a half batch, or step up again to 2.5L for a whole (5G) batch. Half batch then repitch is best, if you have time to brew another batch soon enough.
 
There are a lot of technical considerations when freezing and thawing yeast such as osmotic pressure of solution and speed of freezing and thawing. I have decent wet-lab education, training, and experience which carries over well to the home brewing. I haven't specifically measured my cell counts and viability, but get decent enough fermentation timecourses and flavor profiles in beer.

moz-extension://587cefde-5ecd-4155-bd70-91f54c45d82b/enhanced-reader.html?pdf=https%3A%2F%2Fbrxt.mendeley.com%2Fdocument%2Fcontent%2Ff680aa25-a85f-392f-a227-abc4082552b0

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So, it's likely not as simple as "freezing yeast doesn't work." There will be a heavy influence of the technical aspects of the lab procedures that one implements. I think one key aspect everyone could do is to freeze yeast slowly (about -1C/min) by placing in an insulated container in the freezer and thawing rapidly about about 30-35C.
Sadly those pdf links don't work. Maybe you could upload them directly? I'd love to do a better job keeping yeast alive, I just don't have practical experience so have to rely on what people post and hope it's correct. I'd be glad to learn better. The post I've been following does have the slow freezing/fast thaw info, but I have never heard of the sugar Trehalose, and looks like there's even newer research into improving things: https://www.sciencedirect.com/science/article/abs/pii/S0268005X22006129

Also, my frozen vials have about 5-6ml of slurry. Should this be pitched right into a 1l starter or a smaller one, and 1.040 ok or weaker at first?
 
These ones are straight from a White Labs’ WLP007 pack. ‘Outsourcing’ the lab work, making it incredibly simple. They know what they’re doing. Or make a nice clean starter with your strain(s) of choice and store the same way. The cryovial caps are screwed on tight. They can handle a little pressure, but I haven’t noticed any building up. I recommend just pitching one vial in a 50-100ml first step starter, for simplicity, but I actually just use a loop’s worth to inoculate 10ml sterile wort then step up to 100ml, 500ml, for a half batch, or step up again to 2.5L for a whole (5G) batch. Half batch then repitch is best, if you have time to brew another batch soon enough.
Great, fantastic. Many thanks.
 
These ones are straight from a White Labs’ WLP007 pack. ‘Outsourcing’ the lab work, making it incredibly simple. They know what they’re doing. Or make a nice clean starter with your strain(s) of choice and store the same way. The cryovial caps are screwed on tight. They can handle a little pressure, but I haven’t noticed any building up. I recommend just pitching one vial in a 50-100ml first step starter, for simplicity, but I actually just use a loop’s worth to inoculate 10ml sterile wort then step up to 100ml, 500ml, for a half batch, or step up again to 2.5L for a whole (5G) batch. Half batch then repitch is best, if you have time to brew another batch soon enough.
Sorry, neglected to ask. The aliqouts themselves - they are literally just the Wyeast pack or WL vial, etc., themselves, simply poured into the cryovials - the yeast and wort in the packs, then pitch into 50-100 ml 1st step starter, etc.?
 
Thanks for all of the info on this thread. While I will continue to judge my freezing program, I will also order some 2ml vials and start a parallel fridge program. Just one more step than I usually do from freezing will not be that difficult. (50ml 1.020 to 250ml 1.020 to 2L 10.40)
 
Sorry, neglected to ask. The aliqouts themselves - they are literally just the Wyeast pack or WL vial, etc., themselves, simply poured into the cryovials - the yeast and wort in the packs, then pitch into 50-100 ml 1st step starter, etc.?
Yes, mixed then transferred with a pipette to sterile cryovials. Ideally working in front of a Bunsen burner (or little bleuet burner). Eppendorf tubes are fine too. I just happen to have a box of cryovials hanging around not doing much and I like the more secure screw caps.
 
This thread has got me thinking about my yeast process and handling which is always good. In the area of a portable yeast lab, what does everybody think of one of these to work in? Might seem like overkill but for those of us who have pets it might be an easy way to create a solid workspace and save time with prep and cleanup. Also a CO2 environment could be easily created which would be best for repitching. Thoughts?

https://www.coleparmer.com/p/glove-bag-inflatable-glove-chambers/49332
 
I do my transfers under a propane flame. If my overbuild saved yeast is over a year I use a loop to get the middle portion and have 15 ml screw top tubes of 1.030 wort I pressure can. I do the 15,150,1500 ml steps.
 
Along the lines of the original post, just trying to picture how I can improve my home workspace. As a non-biologist, I am kind of blind to industry practices somewhat. Since I do not have the space to dedicate a clean room or area, what is the next best alternative? Easy and repeatable spring to mind along with compact in size. Sterile seems to = expensive. Where on the sterile vs open air spectrum do us homebrewers need to fall?

I like the repeatability and possible CO2 environment creation of a bag. Filters could probably improve the quality of the inside of the bag compared to the house environment.
 
Since I do not have the space to dedicate a clean room or area, what is the next best alternative?
Low traffic. Low air flow. Sanitize all surfaces.

Not meaning to dis the glove bag. Keeps dust out and dust carries organisms that can contaminate your work. If I was going to fill one with CO2 or N2 I'd probably use a sanitary air filter in the gas line.
 
This thread has got me thinking about my yeast process and handling which is always good. In the area of a portable yeast lab, what does everybody think of one of these to work in? Might seem like overkill but for those of us who have pets it might be an easy way to create a solid workspace and save time with prep and cleanup. Also a CO2 environment could be easily created which would be best for repitching. Thoughts?

https://www.coleparmer.com/p/glove-bag-inflatable-glove-chambers/49332
I’m not sure what that ‘glove box’ is supposed to be used for, tbh, but it isn’t brewer’s microbiology. I don’t care what they claim. It’s not me trying to sell one. The most you’d need, as a home brewer, is a Bunsen burner, which is a lot cheaper and is going to last a lot longer.

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Try not to overthink things. Just be a brewer.
 
Might have missed something but basically you pipette viable yeast into these, store them in the fridge, then do several stepped starters to prep them for use?

The potential problem here, with large volumes of stored slurry, as the slurry ages, is cell counts and assumptions ignore a crucial part of the puzzle, vitality, which declines too over time. This can result in under pitching even if enough cells have been pitched. A case where pitching fewer healthier cells ferments better than more unhealthy cells. By starting at very small volumes, like 10ml, you force things in your favour.

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Might have missed something but basically you pipette viable yeast into these, store them in the fridge, then do several stepped starters to prep them for use?
Yes. Viability seems to remain higher at smaller volumes. Presumably something to do with surface-to-volume ratio and less resistance to nutrient diffusion during storage compared to a whole pack or jar of harvested yeast cake, etc.
 
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