http://confocal-microscopy-list.275.s1.nabble.com/Coverslip-thickness-and-correction-collar-tp591122p591127.html
> Search the CONFOCAL archive at
>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>
> The only way to know which - if either - was the correct answer is to
> stand the coverglass on edge, and measure that. Did you do this?
>
> I did not mention this with respect to the dishes, because that
> measurement would require breaking or unglueing the glass.
>
> Jim P - thanks, thanks, and thanks again, for your point about where
> the lenses are corrected for.
>
>
>
> At 08:52 AM 12/10/2007, you wrote:
>
>> Search the CONFOCAL archive at
>>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>>
>>>
>>> From: Confocal Microscopy List
>>> [mailto:
[hidden email]] On Behalf Of George McNamara
>>> Sent: Sunday, 9 December 2007 6:08 AM
>>> To:
[hidden email]
>>> Subject: Re: Coverslip thickness and correction collar ... option Ib
>>>
>>> Search the CONFOCAL archive at
>>>
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal>>>
>>> option Ib. Measure of the glass thickness before you culture your
>>> cells. Then, for this lens, use dishes whose coverglasses are 170 um
>>> (or whatever thickness works best for the lens at its current
>>> "optimal" setting).
>>>
>>> If one in five are the right thickness, than the cost of that dish,
>>> for this experiment, has gone from about $2 to $10. Should take less
>>> than a minute to measure a dish using a 20x, decent NA, dry lens.
>>
>>
>>
>> Dear George
>>
>> On a recent test I did, I discovered something that I had not
>> expected. A dry objective would significantly underestimate the
>> thickness of a glass coverslip.
>>
>> For a #1 coverslip, a 20x dry objective would measure it as 98microns
>> thick, where as a 63oil objective as 138microns thick.
>> For a #1.5 coverslip the values were 113microns and 160microns
>> respectively.
>>
>> As I said, I wasn't expecting it because the 20 lens was coverslip
>> corrected.
>>
>> So you have to be very careful when using dry objectives to measure
>> anything in Z.
>>
>> regards
>>
>> *********************************
>> Stamatis Pagakis Ph.D.
>> Biological Imaging Unit
>> Biomedical Research Foundation, Academy of Athens
>>
[hidden email]
>>
>>
>>
>>
>>> Compared to the cost of the confocal time (or of an LSM510), or of
>>> your time to do manual image analysis, or cost of the analysis
>>> computer and image analysis software, the extra cost is trivial.
>>> Plus, you can sort the "rejects" and use them with lenses each
>>> thickness is optimized for.
>>
>>
>>> George McNamara, Ph.D.
>>> University of Miami, Miller School of Medicine
>>> Image Core
>>> Miami, FL 33010
>>>
[hidden email]
>>>
[hidden email]
>>> 305-243-8436 office
>>>
http://home.earthlink.net/~pubspectra/>>>
http://home.earthlink.net/~geomcnamara/>>>
http://www.sylvester.org/health_pro/shared_resources/index.asp (see
>>> Analytical Imaging Core Facility)
>>>
>>>
>>>
>>>
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>>
>>
>>
>>
>>
>>
>>
>> George McNamara, Ph.D.
>> University of Miami, Miller School of Medicine
>> Image Core
>> Miami, FL 33010
>>
[hidden email]
>>
[hidden email]
>> 305-243-8436 office
>>
http://home.earthlink.net/~pubspectra/>>
http://home.earthlink.net/~geomcnamara/>>
http://www.sylvester.org/health_pro/shared_resources/index.asp (see
>> Analytical Imaging Core Facility)
>>
>>