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>Hello everyone and hope you all are doing well!
>
>
>I am checking the intensity lost along the z axiso f our lenses 
>which are 40x/.8 water dipping lens and 60x/1.2 w lens;by collecting 
>the z stack from a homogeneous fluorescent sample ,I have noticed 
>the max intensity obtained from 60x lens is dropping faster than the 
>40x dipping lens.I am just wondering this fact is true in general or 
>it's because I'm using a dipping lens?
>
>best
>Sarah


If your homogenous sample is plastic, then it is unlikely to have the 
same RI as the water for which the objectives were designed. Hence 
more SA and more signal loss. In the case of the 1.2 lens, you MUST 
have a coverslip between the objective and the fluorescent specimen.

If the fluorescent specimen is a water solution, then SA is unlikely 
to be the problem and the loss of signal with depth may be related to 
the dye concentration being too high and absorbing the excitation 
beam as it moves into the specimen.

Jim P.

-- 
               **********************************************
Prof. James B. Pawley,               		            Ph.  608-263-3147 
Room 223, Zoology Research Building,               
FAX  608-265-5315
1117 Johnson Ave., Madison, WI, 53706  
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3D Microscopy of Living Cells Course, June 14-26, 2008, UBC, Vancouver Canada
Info: http://www.3dcourse.ubc.ca/	     Applications due by March 15, 2008
	       "If it ain't diffraction, it must be statistics." Anon.