Re: Oil objective maximum area/distance for long-term timelapse imaging

Posted by Sylvie Le Guyader on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Oil-objective-maximum-area-distance-for-long-term-timelapse-imaging-tp7584835p7584836.html

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: [phone]

office: [phone] New number!

LCI website





-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kenneth Chen
Sent: den 2 mars 2016 13:22
To: [hidden email]
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