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There are a couple of points that might bear considering.  One is that, 
depending upon how far the x/y movement is, uneven thickness of the 
coverslip and/or a stage that is not perfectly horizontal will cause one to 
lose focus, especially using high NA objectives.  The other is, if the oil 
is a problem, simply smear the surface you intend to view with oil.  It 
doesn't take much on a surface once the objective is oiled to keep it oiled 
when moving around a surface.
Just a thought.
Carl

Carl A. Boswell, Ph.D.
Molecular and Cellular Biology
University of Arizona
520-954-7053
FAX 520-621-3709
----- Original Message ----- 
From: "Anda Cornea" <[log in to unmask]>
To: <[log in to unmask]>
Sent: Friday, November 17, 2006 12:01 PM
Subject: Re: Multipoint imaging with oil objectives


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Hi Henna,

Is not using an oil objective an option?  Do you have, or could you get, a 
water immersion objective of similar magnification and close NA?  Water 
would be less viscous and would improve spherical aberrations that I expect 
might be an issue when imaging live cells with oil immersion. We use a 
40xNA1.2w to move between 3-5 cells with 0.5-1 minute interval.

Anda Cornea Ph.D.
Head of the Imaging and Morphology Core
Oregon National Primate Research Center
Oregon Health & Science University
(503) 690-5293

>>> [log in to unmask] 11/16/06 11:52 AM >>>
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Dear all,

I was wondering if anyone has done live cell imaging by capturing multiple
points and using OIL objectives. I cannot keep the focus for multiple
points while doing time-lapse using oil. I have tried to stabilize the
system, to pre-heat the oil and to use the objective heater for several
hours prior to imaging. It feels like the oil is not "following" the
objective fast enough, i.e the image is captured before it is in focus. We
have a Zeiss Stallion imaging system equipped with a heating stage, CO2 and
a humidifier. Thank you for your answers in advance!

Cheers,

Henna


Henna Syrjäkari, M.Sc.
Biomedicum Helsinki
Molecular Cancer Biology Program
Haartmaninkatu 8
00290 HELSINKI
FINLAND