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Dear Vladimir,
Yes, but I need to record during the solution changes - on a second by 
second basis - and there is no correction collar....
Greetings,
Christof

On Mon, 18 Mar 2013 12:24:01 -0400, Vladimir Ghukasyan 
<[log in to unmask]> wrote:

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>
>Hello Christof,
>
>As you mentioned, one of the most prominent reasons for the
>distortions you see is change of the refractive index. Lenses are
>usually corrected for chromatic and spherical aberrations for some
>defined refractive index. The more you deviate from it, the more will
>be the extent of the aberrations affecting your image. One way of
>overcoming this is using lenses with adjustable refractive index
>correction collar.
>
>With regards,
>Vladimir
>
>On Mon, Mar 18, 2013 at 11:29 AM, Christof Schwiening <[log in to unmask]> 
wrote:
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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>>
>> Dear All,
>> I wonder if someone could throw some light on a problem we have been 
having
>> for the past few years! We use a Leica SP5 confocal on an upright 
microscope
>> with dipping lenses to image pH and calcium-sensitive fluorescent dyes. 
During
>> solution changes - most notably the addition and removal of organic
>> compounds such as propionate (20 mM)- we get severe image distortion 
and
>> changes in focal depth. I attempted to attach a graph of a ROI from a fixed
>> sealed specimen which we have imaged during solution changes with 3
>> different objectives - however, the ListServer rejected every image format I
>> tried...I can send it by email if anyone is interested. It is impossible that the
>> propionate is actually getting to the specimen itself - I suspect it must be 
as a
>> result of refractive index changes.
>>
>> My questions are: How do others image small structures (i.e. dendritic 
spines
>> or NMJs) during changes of solution with differing refractive indicies? Is 
there
>> some well know trick? Or, am I using the wrong objectives? Does wide field
>> microscopy have the same problems?
>> Many thanks,
>> Christof