From: slk6p@cc.usu.edu
Newsgroups: rec.crafts.brewing
Subject: Re: European Ale (Wyeast 1338) Culture
Date: 31 Aug 92 15:28:55 MDT
Organization: Utah State University

In article <01050166.caj5t2@joebloe.maple-shade.nj.us>, joseph@joebloe.maple-shade.nj.us (Joseph Nathan Hall) writes:
> 
> Hmm.  I have some plates streaked with Wyeast 1338 that look a little
> odd.  The plates are a week or so old now, and there are two sizes
> of yeast colonies on them.  There are a half dozen large (just sub
> BB dia.) colonies on each and many small (ballpoint pen point or smaller
> size) ones.  There are no intermediate sizes.
*Some of the colonies could just be more active than others.  Or they could 
be a contaminant.  I would be inclined to focus on the larger colonies.
Yeast will grow bigger colonies than bacteria (mostly).  Generally you want
that "happiest healthies" yeast bugs.  So for re-culturing, pick the big ones.
*Do not rely solely on size though.  Look at the 
*Do not rely soley on size. Look at the color (creamy white), and the margin
(slightly undulate as they are older, smooth when younger), also look at
their light transmittance, they should be fairly opaque when you look 
through the plate into a light.  
IMHO:  If in doubt- chuck it out.  Start from a fresh starter.  No point
in culturing a mixed culture (unless it's 3056).  Not worth risking the 
beer-  to me.  But see how the subcultures look/smell/etc.  You will see 
variable sizes.

?  I just cultured the wheat yeast (Williams-  a Wyeast product, but is it
3056?  I dunno?)   There do seem to be white, and somewhat darker colonies.
Does anyone know which is which?  I remember that someone successfully isolated
the two types.>

> 
> Evidently the slower-growing (?) strain was more numerous,
> by a factor of 10 or 20, in the starter.
> 
> Meanwhile, I've pitched a brew with the starter, and it seems to be
> progressing normally.  That is to say, it doesn't smell like a hot
> radiator hose or anything, so there must not have been anything 
> drastically "wrong" with the yeast.
> 
> I thought the only double strain Wyeast sold was the wheat beer yeast.
> Contamination of my starter is possible, but certainly it wasn't
> contaminated by anything obnoxious.  I had a B. lambicus culture and
> starter in here a while ago, but I do *not* think that it got loose.
> (I handled that stuff like it was plutonium dust.)
> 
> I just streaked a colony of each size onto another plate and onto
> slants ... stay tuned ...
> 
> Anyone who regularly cultures liquid yeasts is welcome to comment.
> Jack will probably comment anyway.  1/2 :-)
> 
> Can someone out there tell me how to make prepared slides of yeast?
> I've got a friend with a nice microscope and a large set of pH
> indicators (many of which are also stains), and am now feeling the
> itch to scrutinize my cultures at higher magnification....
*First yo
	air dry. It should be free of dust, finger prints, etc.
	2.  Place a small drop of di water (or boiled) on the center.
	3.  Take a small glob of cells with a sterile inoculating loop.
	Mix this into the water to make a paste.  Try to thin it out
	toware the edges. (You want to be able to see single cells)
	4.  Let it air dry.  Then pass it gently over a bunsen burner flame.
	Do not cook it, just heat fix the cells to the slide.  You should
	be able to touch the slide to the back of your hand w/o burning.
	5.  Apply a suitable stain.  Methylene Blue is good.  If you want
	to do differential staining, that's a different story.  If you just
	want to see them...   Flood the slide with stain for ~ 10 -30 sec.
	Then rinse with water to wash off excess stain.  
	6.  Look at the slide under oil immersion.  You should see oval type
	shapes.  Yeast are generally "cocoid" somewhere between rods and cocci.

Hope this helps.  Sorry for the garbage below.  This is a crappy terminal!

			More questions?  E-mail me.
				John Wyllie  SLK6p@cc.usu.edu
dry in the aiw be free of du
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