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Hi Iftach,
We have both types of 2P systems here (an inverted and an upright - both
FV1000 Olympus systems). Our inverted has the standard range of 10, 20
and 40 air objectives and 60 water and oil objectives - the system is
tuned to optimally work with the 60x water. The upright has Olympus' new
25x dipping objective on it.
You will be limited on the inverted to the working distance of your
objective usually before you hit any other limits, our 60 water has a WD
of .23mm (penetration, signal return etc). While the system is tuned and
should work only with the 60x i have had pretty good success with the 40
and 20x objectives as well (the 10x doesn't work at all) and they give
better depth due to the working distance more than anything else.
Obviously the dipping objective gets much better penetration but you are
then limited in what you can do in culture dishes etc. There are ways of
getting dipping objectives working on inverted (i will leave it to Steve
Cody to bring up the self sucking condom technique).
Cheers
Cam
Cameron J. Nowell
Microscopy Manager
Centre for Advanced Microscopy
Ludwig Institute for Cancer Research Melbourne - Parkville Branch
PO Box 2008
Royal Melbourne Hospital
Victoria, 3050
AUSTRALIA
Office: +61 3 9341 3155
Mobile: +61422882700
Fax: +61 3 9341 3104
Facility Website
-----Original Message-----
From: Confocal Microscopy List [mailto:
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On Behalf Of Iftach Nachman
Sent: Friday, 21 October 2011 5:30 AM
To:
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Subject: Re: penetration in 2P inverted vs. upright systems
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These are clusters of differentiating mouse embryonic stem cells (called
embryoid bodies) ranging in size between 100 - 600 um. They need 37C and
5%
CO2 incubation.
Thanks!
Iftach
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