Freezing test tubes of yeast

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Beachbum
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Freezing test tubes of yeast

Post by Beachbum » 7 years ago

A lot of brewers do yeast "slants" in test tubes of a medium like gelatine or agar agar where they culture up yeast and store the tubes in the fridge for future use. I was interested in slanting, to keep a constant supply of yeast and to keep examples of yeast that is often unavailable, for example Wyeast West Yorkshire 1469 that they only release every two years.

Whilst researching slanting, I was presented with the other popular method, actually freezing the yeast and keeping it in the freezer. The problem is that when freezing living matter, the cell walls can be ruptured and kill the cells. This can be avoided in the case of yeast by storing the yeast in a solution of glycerine (glycerol). I have also posted this on other forums as it's of general interest, not solely BIAB.
Image I have an old school freezer that needs de-icing every couple of months so the yeast tubes can be in deep freeze without the interruption that can happen with modern no frost cycling freezers. With modern freezers the yeast can be partly revived during the defrost cycle in the freezer and can have short life, but this can be avoided by storing the frozen yeast tubes in a styrofoam box with a freezer pack to even out the temperature fluctuations.

So I acquired some kit and a bottle of glycerine (to prevent cell walls bursting which is the major no no in yeast freezing). I'll certainly revisit slanting if I get bad results down the track. Also please note I don't have a pressure cooker yet: relying on steaming only.
So here goes: (all measurements in metric, 30ml = about one fluid ounce)

8 test tubes (30ml) to be prepared for freezing. I'm following the method provided by this site:

Into the stockpot:
8 test tubes and caps sitting in a Pyrex bowl,
a few assorted new syringes from the chemist sitting in a couple of Pyrex 250 ml teacups.
Image We are looking to get a cup containing 120ml of 70% water / 30% glycerine, and a cup of 120ml yeast slurry to fill 8 tubes.

Steam for an hour.
Image Arrange a work area away from any spores and yeast as might lurk in your brewery. Using a small graduated syringe, put 85 ml of boiled kettle water and 35 ml glycerine into one of the glass cups or your equivalent bit of kit, and microwave to a boil. The into each test tube, syringe 15 ml of solution, cap the tubes and cool rapidly.
Image Image Image Obtain a cup of slurry / yeast from a starter / topcrop from Yorkshire yeast etc etc.

Into each test tube, syringe 15 ml of yeasty solution. Cap, label and freeze.
Image Image Apparently if you fridge them for 24 hours and then freeze them you get better viability.


I look forward to the results. BTW the yeast is Wyeast 1968 London ESB. B)
Last edited by Beachbum on 02 Dec 2010, 20:25, edited 5 times in total.


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Post by Ralph » 7 years ago

In the usual BB style, what a great pictorial BB! :P

What's the anticipated freezer- life of one of those puppies? I'd guess almost indefinite, i.e. in practical terms, maybe years?
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Post by PistolPatch » 7 years ago

Excellent post BB! You are the man!

I'm looking forward to having a slow, concentrated read of the above. So far, to me at least, it looks like a brilliant "how to" guide. Will look forward to hearing how this goes.

:salute:
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Post by BrickBrewHaus » 7 years ago

Great post!!

When it comes time to use them, do you decant the glycerine/water from the test tube first (I'm assuming the yeast will settle out) to make a starter? Or do you pour the whole thing into your starter wort?


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Post by Beachbum » 7 years ago

When I get to use the first tube I plan to thaw and run the solution off the top (I had them in the fridge overnight and they have settled out with clear liquid on the top) and use the yeast in a starter in much the same way as culturing up a slant, take a couple of days to work up to a pitchable quantity. I think the amount of glycerine left would be minute, but again I might let the starter settle down and again pour the top layer away, then keep stepping up the yeast layer on the bottom of the starter flask.


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Post by outbreak » 7 years ago

Thanks for taking the time to document that! Seems like a good way of saving some money on liquid yeast.

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Post by mordantly » 6 years ago

i have many slants in the fridge from Dec 2009 still! i just built up one of the two WY1056 for my 10gal batch and it was still good. but i make 3-5 slants from a fresh un-smacked pack then pitch remainder into a starter.
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Post by Yeasty » 5 years ago

Bumpity Bump Bump....

Has any one tried this over the last year and a bit ? and what have the result been..I'm thinking of starting to keep slants but this seems a whole lot easier.
Why is everyone talking about "Cheese"

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Post by Aces high » 5 years ago

Here's the Aussie home brewer thread:

http://www.aussiehomebrewer.com/forum/i ... opic=54166

Seems good and I am going to try it with the yeast cake off my black IPA with wyeast 1272. Just need to up the glycerine level as per the AHB thread.

Oops, wrong link... try this one http://www.aussiehomebrewer.com/forum/i ... opic=50154
Last edited by Aces high on 05 Mar 2012, 18:25, edited 5 times in total.

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Post by shibolet » 5 years ago

froze my first viles last night. basically the same protocol as described above. piece of cake as long as everything is squeaky clean.
Cube:
fermenter: Sourdough Spelt Ale, Classic Lambic, Oud Brune, Barrel Aged Belgian Dubbel
Kegs: Bob's Black IPA, Blanc Blond, Soda...
to be brewed:

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Post by shibolet » 5 years ago

update: grew up a vile of frozen WLP550 to a 2 liters starter in a few steps over 6 days. looks very promising right now.
Cube:
fermenter: Sourdough Spelt Ale, Classic Lambic, Oud Brune, Barrel Aged Belgian Dubbel
Kegs: Bob's Black IPA, Blanc Blond, Soda...
to be brewed:


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Post by Philip321 » 1 year ago

Beachbum said
"take a couple of days to work up to a pitchable quantity"

I have a question:- When I come to use one of the vials what exactly is the process. It sounds as if you pitch the vial into a starter let it do its thing and then start all over again to increase the quantity. Is this correct. Do you put the starter into the fridge after the first ferment or do you just keep adding DME?
I'm really confused.

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Post by Lumpy5oh » 1 year ago

You make a starter and then let the yeast settle or cold crash in the fridge. Once settled decant the old wort off of the yeast and replace with fresh wort and ferment again. Keep doing this until you get what you need.
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Post by Philip321 » 1 year ago

Thanx for the info Lumpy5oh.
So you make a starter let it ferment for a couple of days then cold crash it in the fridge. When do i know it has fermented enough?

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Freezing test tubes of yeast

Post by Lumpy5oh » 1 year ago

24 to 48hrs is enough for a starter.
Here is a link to White Labs Starter help.
http://www.whitelabs.com/resources/home ... arter-tips
Last edited by Lumpy5oh on 19 Dec 2015, 08:45, edited 5 times in total.
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Post by Philip321 » 1 year ago

Would these plastic test tubes/vials and syringes be able to stand the temperature in a pressure cooker?


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Post by joshua » 1 year ago

Phillip, Why do you want to use a pressure Cooker??

The plastic tube may melt at 164F or 250F depending on the Plastic used.

There are many ways to Sterilize with a Long soaking in Sodium metabisulfite,
see(http://www.haurakihomebrew.co.nz/cleani ... phite.html

97% Alcohol does a good Job, but, will mess up the Plastic.
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Post by Philip321 » 1 year ago

Thanx for the reply Joshua,
I've read quite a lot on Yeast harvesting and freezing the yeast and they all sanitise using boiling water and most use a pressure cooker.

I would prefer to sanitise using a sanitising solution as you mentioned.

So you think it will be ok to use a sanitising solution and not boil?


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Post by joshua » 1 year ago

Yes, if the The Tubes are clean and Spotless, the Sanitizing solution will sanitize in a few Seconds/Minutes, and Sterilize in and Hour.

The Sodium Meta-bisulfite destroys all bacteria, viruses, Molds, and Fungus.
After it is dry is safe for brewing yeast.

Boiling at 100C it will take up to 6 hours the Sterilize most things.
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Post by mally » 1 year ago

joshua wrote:Boiling at 100C it will take up to 6 hours the Sterilize most things.
Just FYI - this is why pressure cookers are employed. 30 mins at 121C (standard pressure cooker) or 4 minutes at 132C (autoclave).
Last edited by mally on 23 Mar 2016, 20:49, edited 5 times in total.
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Post by PistolPatch » 1 year ago

Philip321 wrote:So you think it will be ok to use a sanitising solution and not boil?
No.

For several reasons...

Any chemical you use must be a 'no-rinse' one.

Any 'no-rinse' sanitiser I know of will not render something sterile, and, from what I know, this is one area of brewing you want to be as sterile as you can get. (Note that I have never done the procedure myself).

I am not aware of any 'no-rinse' steriliser. That's dodgy advice from the HBS that Josh linked (and I have no idea where the one hour thing came from). I used to use that chemical, years ago, when working for Matilda Bay. It's an awful chemical to use and it's definitely not a no-rinse steriliser. I wouldn't even use it as a no-rinse sanitiser.

...

Philip, in your post, you also asked if "these plastic test tubes/vials and syringes [meaning the ones in the original post]" would be able to stand the temperature of a pressure cooker.

I think that is an impossible and unsafe question to answer here. I'm not even sure that the plastic equipment used in the original post is even graded to handle 100°C/220°F let alone temp of a pressure cooker which can get up around 120°C/250°F.

Some plastics might not melt at certain temperatures but will give off certain tainting chemicals.

For a good answer on this, I think you would have to ask the supplier, before you buy, and get the answer in writing (seriously).

Unless someone else jumps in with a definitive answer, do the research and please let us know how you go.

:peace:
PP
Last edited by PistolPatch on 23 Mar 2016, 21:31, edited 5 times in total.
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Freezing test tubes of yeast

Post by Lumpy5oh » 1 year ago

I would think these are a better option.
https://www.amazon.ca/gp/aw/d/B004RYF8K ... 9S88LI0VOY
Last edited by Lumpy5oh on 29 Mar 2016, 21:41, edited 5 times in total.
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