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Can I get some help with my 1st yeast starter, please?

Silicon

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I'm trying to plan a yeast starter for the 1st time, making a Marzen, OG 1.050, 5gal batch, Wyeast yeast

Someone at American Homebrewers Association suggest a couple online calculators, which I tried, plus added Mr. Malty, but they all gave different results:
Brewer's Friend says 2L starter yields 419B cells
Brew United says 2L starter yields 402B cells
Mr. Malty says to get 350B, I need a 2.74L starter

I was looking at a White Labs video* and they have an Inoculation Rate Graph which says for 2L starter volume, inoculation rate 50, total cells at finish is 205B. If I'm reading their graph right, I figured I'd need a 2-stage starter, 1.5L each. I'd think I could trust White Labs more than those online calculators, so maybe I'm just not reading their chart right? WL's standard starter is a 1L starter for ales, 2L for std lagers, so that seems closer to those calculators.

I've read that there's some sort of law of diminishing returns with making bigger and bigger 1-stage starters which, I think, is the whole point of going to multi-stage starters. A 2L starter doesn't sound all that big, so the chart is now confusing me.

Can anyone else shed some light on this, please?  Man, I need a beer!

* White Labs video: https://www.youtube.com/watch?v=1zUYxb-_B8A (6:12 mins into video)

PS I'm attaching a screenshot of WL's chart that's 6:12 mins into their video.
 

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Don't expect a consistent answer from any of the calculators.  Each has been created using one of usually 4 or 5 sets of data, most of which are independently collected.  Depending upon the care and effort put in, the quality of the experimentation conducted, and the skill of the person doing the cell counts, the data is more or less reliable.

Having said that, without verification of cell counts, all of them are pretty much a SWAG at what anyone else would get in their starters.  I find that the actual number of cells produced from a starter varies quite a bit based upon the strain and temperature the wort is held at. 

The other thing to keep in mind is that the goal is to produce healthy cells with thick strong cell walls.  These are capable of rapid reproduction when introduced to the wort without growing too weak to finish fermentation.  The actual count of cells is almost secondary to this goal.  There is also a pretty wide target when it comes to pitching rates.  You can easily undershoot and have issues, but rarely can you produce so many cells that you sacrifice yeast flavors (both good and bad) from a single starter in a standard batch.  Pitching a wort on top of a full yeast trub from the previous brew is one way to push at over pitching.

So going back to your brew, you have a Marzen at 1.050 with 5 gallons in the fermenter.  According to BeerSmith you will need approximately 352 billion cells.  Depending upon the age of your package, you will probably need to do at least the 2.74 L starter.  You can step this up in two steps, but I would recommend making the first starter and then splitting the yeast slurry for the second.  If the cells are too crowded in the second starter they will not produce as much as when they have plenty of growing room and more food per cell.  You don't want to grossly under pitch the starter because the cells can only reproduce so many times before their cell walls are too thin to do much more. 

If I were tackling this as a one-off with the yeast, I would do back to back 1.6L starters in a 2-L flask on the stir plate or split the packet between two 2-L flasks and do the same size starter in each.  This should give you plenty of cells to pitch without creating a gross over pitch.  I tend to pitch a bit on the high side with lagers anyway. 

Which Wyeast strain are you using?
 
Not only are you a great help, but you're also a mind reader. I was thinking of doing 1.5L for a 1st stage, then split it equally into a pair of 1.5L 2nd stage starters.  That was my initial gut feeling based on what I got out of White Labs' data combined with what I read on Wyeast's site.  I figured it might make a good base starting point for lagers. Then, for normal ales, either a single 2L or a 1.5L with a 2nd 1.5L 2nd stage, depending on the recipe's OG. (Not considering another tripel for a while, that batch was a real beast!)

The back-to-back 1.6L method you mention...are you thinking of a standard 2-stage with 1.6L each, decanting the 1st starter and adding 1.6L for the 2nd stage?

So thank you tremendously, I really appreciate it! :)

Oh, and I'm using a Wyeast 2124 Bohemian Lager yeast. My 1st Marzen used a Wyeast 2206 Bavarian Lager yeast, but I used a 2124 in my last Marzen and preferred it just a tad more. I'm not sure which I'll eventually end up with as my technique still has too many variants in it, which is what Batch 14 is all about...I'm trying to go more "science" and less "art" this time. ;) It's my 1st time using a yeast starter, but I think that'll have the greatest impact on consistency since the rest of my processes are pretty stable by now.

PS I've also been curious as to seeing what would result if I started with a 0.5L starter, wait for fermentation to peak, then start slowly adding 2L of wort at a slow, steady drip over the course of a full day, the idea being trying to keep the yeast in as constant peak aerobic state as I can. Then, if it works, I'd try more wort and longer periods and chart the comparisons. But first, I need to save up for the microscope, etc. (My budget is hardly big enough to call it a budget, lol...but that's ok, slow 'n steady is fine...I teach patience to rocks.)
 
Huh. Dunno why I never thought of it, but looked at BeerSmith's starter tool for the 1st time. (Yeah, go figure, duh!)  Target is 350B: 1.78L starter, 6.69oz DME, 1.040 OG. Would that be a single stage starter? I don't see how to use the tool to do a 2-stage starter, though...maybe in version 3?  I'm still using v2 since I won't upgrade because of the change in licensing.
 
BeerSmith 3 can do 2-stage yeast starters, BeerSmith 2 can only do single-stage. I find that the BeerSmith 3 calculations are very close to the Homebrew Dad's online calculator. My skepticism regarding accuracy is exactly the opposite of yours. I trust the online calculators more than the White Labs web site. I made my first yeast starters using the White Labs instructions but eventually found out that their instructions are (or were) totally messed up. Unless they have updated them since I first tried them, the instructions at https://www.whitelabs.com/resources/homebrew-starter-tips have DME quantities listed that are totally wrong. The online calculators may disagree on some things, but they will all give you accurate information on how much DME to use to get a gravity of 1.035 - 1.040.

--GF
 
a single 2L starter with Fresh (check date) yeast should easily make you 350-400b cells on a stir plate, especially if you dose a tiny bit of O2
 
Having just looked at the White Labs information that Silicon referenced, it is really a dumbed down version of a yeast starter calculator designed more for people not to have to go through the data entry of the actual model.  Like all the other calculators, it is probably accurate for the data set they produced.  Being yeast manufacturers, they most likely get near the maximum reproduction from their starters.  Heck, they know yeast!

WRT starting with a 500 ml starter and then adding more, my inclination is that this would be a nice wake up for the yeast from cold conditions, but I am not sure that you will get much reproduction from that first 500 ml and limit production from the addition of the remaining wort.  Without actually doing this and conducting cell counts, I have nothing concrete to base this on and am just projecting from past efforts with small starters.

I have used WY2206, WY2308, and WY2124 in the past years brewing a Marzen.  The best one that I brewed was with WY2633, but my LHBS only carries that as a special order and I use the 2206 and 2124 for most other lagers.  So I usually bounce off of these and keep a culture going of each strain and rotating them throughout the lager brewing period.  Of the two I have used most often, the 2206 seems to be a bit more malty.  I gave up on the 2308 as I needed to watch it more carefully for a diacetyl rest.

Reliability is one reason that I dragged out my microscope and bought the supplies needed to do cell counting.  For the past two years, I have tracked pitching rates of viable cells to give me more consistent results.  I never had too much of an issue with lagers as I tended to pitch a bit higher than 1.5 Mil/ml/Plato.  Ales were another matter, especially English styles.



 
I also notice that the numbers provided in that video contradicts published information from Chris White.

so that's interesting....

as far as pitching to a 500ml starter, I wouldn't recommend it, as it may cause the yeast more stress than benefit considering its not really enough food to get them going. ideally you should try to pitch in the 50-100m cells/ml range.

I can't recommend the "yeast" book enough. It's written by Chris and Jamil, two of the leaders in the yeast world right now.
 
GigaFemto said:
BeerSmith 3 can do 2-stage yeast starters, BeerSmith 2 can only do single-stage. I find that the BeerSmith 3 calculations are very close to the Homebrew Dad's online calculator. My skepticism regarding accuracy is exactly the opposite of yours. I trust the online calculators more than the White Labs web site. I made my first yeast starters using the White Labs instructions but eventually found out that their instructions are (or were) totally messed up. Unless they have updated them since I first tried them, the instructions at https://www.whitelabs.com/resources/homebrew-starter-tips have DME quantities listed that are totally wrong. The online calculators may disagree on some things, but they will all give you accurate information on how much DME to use to get a gravity of 1.035 - 1.040.
Isn't it 10:1 water to DME?

Aye, the Yeast book's on my wish list, will have to buy if if the kids don't get it for my birthday. :)  So far, everything else has been easy, but as I found out with baking artisan breads, yeast is really complex, not so straight forwards as it'd seem.

So I've got a WY2124 atm, curious if I give the WY2206 another go, though, esp. now that I'm learning to get pitching right...that should give me much more consistent batches. It seems like my prior batch comparisons wouldn't even be that valid, seeing how much of a difference getting the whole yeast thing down pat really is.

So sounds like the general consensus is that a straight-up 2L starter will do perfect for a 1.050 Marzen using pretty fresh yeast.

1 last question: if I do a 5g batch, then harvest the yeast and immediately do another identical batch, is there a general rule I could follow as to how much of the yeast I should reuse? I know it won't be as lively as the 1st batch, but others are saying that the cell count increases by 5x or 6x, so I assume I'd be using just a portion of it? Half, maybe? A quarter? (That's just a SWAG.)
 
Silicon said:
1 last question: if I do a 5g batch, then harvest the yeast and immediately do another identical batch, is there a general rule I could follow as to how much of the yeast I should reuse? I know it won't be as lively as the 1st batch, but others are saying that the cell count increases by 5x or 6x, so I assume I'd be using just a portion of it? Half, maybe? A quarter? (That's just a SWAG.)

There are a variety of ways people reuse the yeast at the bottom of the fermenter.  The first, easiest, and laziest is to just pour your next batch directly on top of the trub left at the bottom of the fermenter.  While this would most likely be a significant over pitch, you are also carrying over an extra ripe cake of trub which is not yeast or composed of dead yeast cells.  It works for a repitch or maybe two, but sooner or later you will start to build up other off-flavors from the decaying trub and dead yeast cell autolysis, not to mention that other stray bacteria colonies would have a field day on the left over cake.

The second is to leave behind a fair amount of beer and use that to slurry up the yeast cake.  Allow it some time (30 minutes to an hour depending upon the flocculation quality of the yeast) to settle and pour off the suspended yeast/beer slurry into sterilized or sanitized pint or quart jars.  You can then refrigerate the jars and collect the yeast cake on the bottom.  This will be a combination of yeast and light trub and should be white to cream in color.  When you have a settle yeast cake you can estimate the concentration of yeast and density of the cake by assuming that the compacted yeast has a count of approximately 4.5 billion cells per ml.  Depending upon how much trub you have mixed in, you can estimate a percentage of yeast to trub and discount this figure by your estimate to get close to a reasonable amount per ml of cells.  Multiply this out by what you need to pitch and you have the volume you should measure out for your next batch. The beer in the solution should prevent other contaminants from growing.  I would guess from my notes that this amounts to approximately half of what I collect in a following batch, though much depends upon my patience and available time which determines the cleanliness of the collected yeast cake.

The third is similar to the second except that instead of leaving a fair amount of beer behind, add a quart of sterilized water to slurry up the yeast/trub into.  Proceed as above.  This is my usual means of re-use of the yeast.  I find that the resultant yeast cake is cleaner and brighter and healthier.  The fluid is always diluted beer, so there is some alcohol present as a mild preservative though not enough to prevent issues if you carry over any contaminants from the previous batches or collected during the transfers.

Another method that I also use is to over build the starters.  Since I do cell counts, I can remove from the starter the amount that I need for a batch and then either store or propagate more from the remaining yeast which is left over.  This is becoming more popular as you don't have to worry so much about any character from the previous batch carrying over to the next batch.  People feel that there is less chance of contamination, though I have not seen any difference in my efforts. 

Overall, I use a combination of the last two methods described.  I find that I get better results from harvesting most of the ale strains and then building up with starters once I get to the third or fourth generation (repitch) as that generation is where most of the strains I use really hits its stride.  I've gone as many as 9 generations, but have found too many off-quality cells and less uniformity in cell appearance.  I will also do a culture plate of strains that I intend to use again later in the same brewing season.  This has helped me keep strains that I start with in the fall to reuse in the spring surrounding my run of lagers.  It also starts me off with clean colonies of yeasts without worrying too much about contaminants.  In the 4 years I have been doing this, I only came across one strain that showed any significant contamination and needed to be tossed, so I count myself lucky in that regard.

Lager strains (including hybrids such as WY2112) are a bit different.  Here constant harvesting and repitching leads to issues with flocculation and settling.  So I start with splitting small batches between two strains (usually a Helles or festbeir type of recipe) to grow up a base of yeast. I can repitch or propagate from those initial collections to fill in the next 10 to 12 brews by alternating the use of the two strains.  Since I generally want my lagers to be clean and crisp, working from this initial harvest and using starters from that point to propagate further seems to keep the risk of mutations and off-flavors showing up.

Such reuse requires planning.  I will use 8 different strains this year for approximately 34 to 36 different brews.  Between the cost of DME, yeast nutrient, and initial cost of the yeast packs, I estimate that my yeast cost runs to just over $3/brew versus $7 a pack.  This does include a few one-offs (such as WY1007 for an Altbier and WY2298 for a Czech Pilsner) so that tilts the picture a bit on the number of strains versus reuse. 




 
pretty good tutorial on yeast washing on the brewers friend page.

https://www.brewersfriend.com/2010/01/30/yeast-washing-101/

personally i've been starting from plates lately, so I'm no help ;) i don't really prefer to re-pitch as I've got irrational concerns about excessive mutations. i make my plates and slants (5 slants, 20 plates) from a fresh pack of yeast, and bring up brewing yeast from there.

I could easily reduce the number of plates i make as i typically end up tossing a few... its a work in progress to figure out a work flow.
 
dtapke said:
pretty good tutorial on yeast washing on the brewers friend page.

https://www.brewersfriend.com/2010/01/30/yeast-washing-101/

personally i've been starting from plates lately, so I'm no help ;) i don't really prefer to re-pitch as I've got irrational concerns about excessive mutations. i make my plates and slants (5 slants, 20 plates) from a fresh pack of yeast, and bring up brewing yeast from there.

I could easily reduce the number of plates i make as i typically end up tossing a few... its a work in progress to figure out a work flow.

I have to be honest.  I cringe when I hear the term yeast 'washing' as it really is misused in common practice.

Yeast "washing" is rinsing the yeast slurry with an acidified water wash, generally using phosphoric acid.  The acid content of the slurry should be between 2.0 and 2.5 to reduce the amount of any contaminants which may be present, such as lactobacillus-type bacteria, wild yeasts, or mold spores.  To work effectively, the slurry must be stirred for at least an hour under the acidic conditions.

What most people actually practice is yeast 'rinsing' which is the act of trying to reduce the amount of trub and non-yeast material in the slurry.  The goal here is to end up with mostly live yeast cells with little other trub coming along for the ride.

I hear you about plates.  When I was saving all my yeast strains through making culture plates, I came to accept that I will make about 2 to 3 times the number of plates than I will use.  Then I would need to propagate a few colonies from each plate every 6 to 8 months just to preserve that strain.  It got to be enough of a time suck, that I started using the plates only to 'clean up' those yeast strains that I needed to preserve for a few months.  I keep about 4 strains from Spring to Fall and then again from early Winter to Spring using cell plates at this time.  Makes the job easier and has been a real help in determining if I start getting sloppy with my sanitization at any point.
 
LOL! i hear you, I've honestly never done it but am familiar with the terms and concept and you're totally right, but for the layman who's learning the link is a pretty decent resource to helping out with rinsing.

as far as long term storage, I've recently bought a microfuge to spin yeast out and then store in the freezer, my deep freeze runs -10 which should suffice for 2-3 year storage suspended in glycerin. I figure i can store 50 or so 2.5ml vials of each strain, and then pretty quickly just run one of those up to a full pitch when brewing with that strain. I'd like to bank closer to 20 strains this way.

keeping slants in oil helps reduce the need propagate for preservation, which is why i've been keeping a few that way, then when the plates get old i just toss em.

It's an odd hobby inside of a hobby, thats for sure!
 
I never had a lot of luck with freezing yeasts.  It was early in my brewing, so that may account for some of the issues.  I've learned a lot about microbiology since then.  Besides, since I cracked the seal on my semen tank I don't have the deep freeze capability anymore.

Lately, I've been looking into isotonic storage of yeast and doing much reading on that technique.  Since I have access to everything I need at work or at home, I am interested in trying it out. 

When I first read your list, I thought '20 yeasts, holy crap!'  Then I opened up my inventory tracking spreadsheet and I have 25 yeasts listed there.  Only about 14 are common use with others being for specific recipes.

Another link which explains yeast washing and rinsing well is https://www.homebrewersassociation.org/how-to-brew/yeast-washing-yeast-rinsing-whats-difference/ from the AHA. 
 
Oginme said:
There are a variety of ways...

The second is to leave behind a fair amount of beer...

The third is similar to the second except that instead of leaving a fair amount of beer behind, add a quart of sterilized water...

Another method that I also use is to over build the starters.

I'm leaning strongly towards #3. I especially like the diluted beer aspect, makes a lot of sense to me. My gut says this is a good base I can start with, then branch out from there and experiment. I've been having lots of ideas this week, but I think I'll play it safe, start slow, and wait for experimentation once I get and read the Yeast book. Many thanks for your thoughts and experience, I mean it. The more I read online, the more varied everyone's opinions and experiences get, and I'm seeing a lot of conflicting statements from people, making it hard to come up with a good plan. But, what I've been getting from you guys, esp. you, dtapke, and brewfun jibes really well and makes lots of sense. I know brewing is half science and half art, but it seems elsewhere is slanting more heavily on the art side of things. ;)
 
Oginme said:
personally i've been starting from plates lately...

Plates?

Oginme said:
Yeast "washing" is rinsing the yeast slurry with an acidified water wash...

What most people actually practice is yeast 'rinsing' which is the act of trying to reduce the amount of trub and non-yeast material in the slurry.  The goal here is to end up with mostly live yeast cells with little other trub coming along for the ride.

I didn't know of the 2 different terms. I've been saying "washing" when what I mean is "rinsing", so thx. :)
 
Silicon said:
I'm leaning strongly towards #3. I especially like the diluted beer aspect, makes a lot of sense to me. My gut says this is a good base I can start with, then branch out from there and experiment. I've been having lots of ideas this week, but I think I'll play it safe, start slow, and wait for experimentation once I get and read the Yeast book. Many thanks for your thoughts and experience, I mean it. The more I read online, the more varied everyone's opinions and experiences get, and I'm seeing a lot of conflicting statements from people, making it hard to come up with a good plan. But, what I've been getting from you guys, esp. you, dtapke, and brewfun jibes really well and makes lots of sense. I know brewing is half science and half art, but it seems elsewhere is slanting more heavily on the art side of things. ;)

You will need a good supply of sterilized water.  I use quart canning jars and a pressure cooker to sterilize both the water and the storage jar.  I can do up to 7 jars at a time and always keep some handy for other uses, such as topping off a fermenter, rinsing flasks of iodophor (when I don't have the time to allow it to dry naturally), quick starters, etc. 

As far as the art/scientific balance, there is a lot of anecdotal information around when it comes to home brewing, along with myths and old wives tales.  I try to preface my observations as just that or as my experience when I am not completely sure of the universal applicability of my practice.  I always like to divide it by process which is science and recipe which is an art.  There are cross-overs to be sure, but the general concept holds. 

I count myself lucky in that I work in a science based field with a lot of very intelligent people around me.  This also gives me access to a lot of publications and data sources which are not available to the general public so I will sometimes go on binges of gathering papers and articles about a given aspect of brewing to get to know the subject matter as well as I want. 
 
Silicon said:
Oginme said:
personally i've been starting from plates lately...

Plates?

By plates I am referring to cell culture plates or petri dishes.  It is an alternative to slants which are done in cuvets or tubes and tends to take up more space.  At my age, I find the open plates easier to deal with than the narrow openings on slants.  I can easily see the different colonies and pick off the ones that I want to reproduce more readily using the plates.
 
Art is inherent in science, and generally vice versa.

Yeast plating/slanting is a process where you start with a petri dish, fill it with a media (malt agar) and then deposit literally the tiniest amount of yeast (ideally single cells) across it so that they can create colonies. This allows you to start multiple yeast starters from "clean" cultures. It's really, really, nerdy. its like a hobby within a hobby.

For me, i do it because i brew small nearly commercial size batches (15-30g) and it allows me the ability to get a lot of healthy, mutation free yeast for really cheap. Re-Pitching (washing/rinsing) is one way of saving $ on yeast, and also reusing which is great, unfortunately it has its downsides. Mutations, breeding higher flocculatting yeast, higher infection chances, and others.

I'm not saying re-pitching is bad, it's just not for me. As they say, "Your Mileage May Vary"

my practice will be different than your practice will be different than oginmes practice will be different from Vinnie Cilurzos will be different than (keep going....)

you've gotta develop your own workflow and habits. It's great to pull from the resources and learn from other brewers, but you've also got to find your own path.
 
Oginme said:
You will need a good supply of sterilized water.

I count myself lucky in that I work in a science based field with a lot of very intelligent people around me.  This also gives me access to a lot of publications...

ditto. I'm a sr. software engineer at the major aerospace company in WA.(Systems Design & Integration Specialist, now doesn't that sound fancy? lol) We have full text for all journals, articles, etc. and I know how to find anything...I once dev'd for Serials Solutions. :D) Anything I want, any time, and I don't even have to leave my desk. Lots of engineering types around, not many homebrewers, though. (I'm working on that...)

I'm also kinda lucky in that our water profile is really close to my German beers, and not too far off for my Scottish ales, either. My water goes through 2 filtration systems, then boiled, and stored--short term only--in a glass carboy. (I use it for my sinus rinse stuff, too, hehe.) Even my sourdough starters have never been invaded by bacteria. (Pineapple juice ftw!) That's what got me started on my yeast learning.

dtapke said:
Art is inherent in science, and generally vice versa.

Yeast plating/slanting is a process where you start with a petri dish...

I totally agree about the art+science part, but I think most people don't get it. Everything's inter-related and I have to be fairly good at everything, goal's to be a SME for the most important things. I'm an inventor by nature and the holism, I believe, is what allows me to look into rooms through windows others don't use. My talent is analyzing things & processes, storing it away in my mental toolbox, then--at really weird times--these processes pop into my head, giving me some totally different way of doing something new. I can see everything working w/o closing my eyes. (The kids think it's funny.) Sometimes, someone's thought of it before me, and that's cool. But sometimes, it's something unique, and sometimes even revolutionary or, at least, evolutionary. I'm also a perfectionist, though, which is why I know I'll eventually be doing petri dishes, simply because now I know that I can. :)

Some day, I'll be where you are, or at least close. Kids in school, brewing out of the kitchen, garage being used for storage, concrete foundation has an RV on it instead of a workshop...but that'll change in a few years. Meanwhile, it's low-budget kitchen brewing for me. I'm patient, though.

Ok, now off to start my 1st starter. (Man, doesn't that sound redundant?)  The plan is a plain 2L starter, rinse a 1.050 Marzen's trub after moving it into the secondary, then immediately use all the yeast in a 2nd 1.065 Marzen where I'll need the extra yeast, thus the whole thing instead of 1/2.

Thx again guys, I'll let you know how this turns out next week. /cheer
 
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