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I was told at SFN that one of the reasons for silicon immersion oil, is that glycerol changes its RI during prolonged imaging sessions.  Anyone know who much or how soon?

If it is significant, it is a good argument for silicon corrected lens - especially if doing long 3D reconstructions of thick tissue.

Mike Ignatius, 
Molecular Probes/LifeTechnology

No commercial interest.


-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Lloyd Donaldson
Sent: Monday, January 10, 2011 11:22 AM
To: [log in to unmask]
Subject: Re: Glycerol Objectives - experience with

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Jeremy

We have a Leica 63x glycerol objective with adjusting ring which gives very noticeable improvement over 63x oil which we also have. We also have a 20x multi immersion objective that can use glycerol. We have both oil and glycerol objectives, although we prefer glycerol as a mounting medium it is incompatible with some of our samples because of induced swelling so we have to mount in oil for those samples. We also have the oil lens because it is UV compatible - we have a 355 nm laser.


Dr Lloyd Donaldson

Senior Scientist, Project Leader - Microscopy/Wood Identification
Scion - Next Generation Biomaterials
Private Bag 3020, Rotorua
New Zealand 3010

Ph: 64 7 343 5581



-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Jeremy Adler
Sent: Monday, 10 January 2011 9:39 p.m.
To: [log in to unmask]
Subject: Glycerol Objectives - experience with

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We are considering buying a new confocal for examining fixed and
potentially thick biological specimens.
Given the need to have to an RI match between the specimen, mounting
medium and immersion medium, I want to ask about the pros and cons of
glycerol objectives.

Oil objectives appear to have no obvious advantages, the higher RI
will only apply if specimens are mounted in a medium with a RI close
to that of oil - while most media seem to have lower RIs. Why would
anyone choose an oil objective  and which mounting media work for
thick specimens ?

A possible disadvantage of glycerol objectives is that glycerol is
hygroscopic which could change its RI. Are there any RI equivalent
immersion media ?.

So the pros and cons of oil or glycerol objectives.







Jeremy Adler
Genetics & Pathology
Rudbeckslaboratoriet
Daghammersköljdsväg 20
751 85 Uppsala
Sweden

0046 (0)18 471 4607

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