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We regularly image the 4 center wells (about 50 total points) of a Labtek 8
well chambered coverslip for 8-16 hours at 37c using PFS on a TiE.

First we put the plate on, one drop of oil on the 60X or 100X objective,
and drive to all four wells.

Then we take the plate off, and add one more drop of oil, and we're good to
go.

We also slow the stage speed way down so PFS can follow the glass.

Hope this helps.

Gary

On Wed, Mar 2, 2016 at 7:49 AM, Sylvie Le Guyader <[log in to unmask]>
wrote:

> Hi Ken
>
>
>
> My experience: imaging 2 cells 1.7mm apart (thickness of the well wall in
> the MW plates we were using then) every 10 min for 12h with a 60x oil
> objective and a hardware autofocus at 37 deg resulted in a high risk (about
> 25% of the cases?) of losing focus. There go years of sweat and tears…
>
>
>
> We were lucky to get to test a super device manufactured by EMBLEM, the
> commercial side of EMBL. You send your objective. They custom make a cap
> and send you a kit containing cap, tubes, pump and instructions. It is a
> bit clumsy because you need to remove all other objectives and you need to
> be careful not to put oil all over the place but it works a charm. I have
> imaged 10 positions in each well over 15 wells every 10 min for 15 h with
> this device without losing focus. :) You could contact them<
> http://www.embl-em.de/articles.php?lang=de&hid=4&Cat=Technology_Offers>
> to ask if they commercialize it.
>
>
>
> Med vänlig hälsning / Best regards
>
>
>
> Sylvie
>
>
>
> @@@@@@@@@@@@@@@@@@@@@@@@
>
> Sylvie Le Guyader, PhD
>
> Live Cell Imaging Facility Manager
>
> Karolinska Institutet- Bionut Dpt
>
> Hälsovägen 7,
>
> Novum, G lift, floor 6
>
> 14157 Huddinge
>
> Sweden
>
> mobile: +46 (0) 73 733 5008
>
> office: +46 (0) 08-524 811 72 New number!
>
> LCI website
>
>
>
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[log in to unmask]]
> On Behalf Of Kenneth Chen
> Sent: den 2 mars 2016 13:22
> To: [log in to unmask]
> Subject: Oil objective maximum area/distance for long-term timelapse
> imaging
>
>
>
> *****
>
> To join, leave or search the confocal microscopy listserv, go to:
>
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>
> Post images on http://www.imgur.com and include the link in your posting.
>
> *****
>
>
>
> Hi all,
>
>
>
> We're in the process of designing microfluidic devices for multi-day
> time-lapse imaging. In the past we've had some trouble with image quality
> for very large devices deteriorating due to the loss/thinness of the
> immersion oil layer between sample and objective when very large areas are
> scanned. We have only used traditional oil objectives (no water or
>
> glycerol) Does anyone have any strategies to mitigate this or rules of
> thumb about "safe" maximum sampling areas or distances to travel?
>
>
>
> Thanks,
>
> Ken Chen
>



-- 
Best,

Gary Laevsky, Ph.D.
Confocal Imaging Facility Manager
Dept. of Molecular Biology
Washington Rd.
Princeton University
Princeton, New Jersey, 08544-1014
(O) 609 258 5432
(C) 508 507 1310