micromanipulation of veg. cells

Neigeborn, Lenore neigeborn at OCELOT.RUTGERS.EDU
Mon Aug 5 13:15:17 EST 1996


Hi Kevin,

What strain background are you in?  We've found that the ability to separate and
timing of separation of mother and daughter are very strain dependent.  We have
no trouble with W303.  On the other hand, with SK1 we don't get separation of
the first mother and daughter until there are a total of 4 full sized cells
(whichm, as you know makes it difficult to tell who is mother and who is
daughter)!

We've considered applying some glusulase to the area to see if we can promote
separation, but we're not quite sure how to go about it without glusulasing the
cells to death.  Don't the pombe people glusulase their tetrads in situ prior to
dissection?  Something like that might help.

Lenore Neigeborn
Waksman Institute
Rutgers, the State University of New Jersey
PO Box 759
Piscataway, New Jersey  08855

ph:  (908) 445-3668
FAX: (908) 445-5735
email:  neigeborn at mbcl.rutgers.edu
_______________________________________________________________________________
To: yeast at net.bio.net
From: Kevin A. Morano on Mon, Aug 5, 1996 2:03 PM
Subject: micromanipulation of veg. cells
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Date: Mon, 05 Aug 1996 11:07:12 -0500
From: kmorano at umich.edu (Kevin A. Morano)
Subject: micromanipulation of veg. cells
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Hi all,

I'm trying to manipulate vegetative cells using a sick Rainin apparatus
and am having trouble separating daughter and mother cells.  I am doing
all of this on an inverted YPD plate and can move the cells around no
problem.  The hangup occurs when I try to separate a large bud from mom or
worse, when the bud fails to pinch off before a new bud emerges.  For my
analysis I really need to be able to recognize who budded from whom and
get them away ASAP.  Does anyone have advice on encouraging daughter yeast
to move out?  WHile I'm at it, does anyone know why they stay clumped for
so long and continuously make new buds before separating?

Thanks in advance for any advice!

Kevin





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