Re: Implications of no coverslip on objectives corrected for coverslips
- From: renevanwezel@xxxxxxxxxxx
- Date: Fri, 6 Feb 2009 02:12:10 -0800 (PST)
On 4 feb, 00:54, Ian Shieh <ish...@xxxxxxxxxxxxxx> wrote:
Hi,
I am using a Nikon C1 confocal microscope to image surfactant molecules
at an air-water interface. Currently, I use a Plan Apo 20x objective,
NA = 0.75, air "immersion" with 0.17 mm coverslip correction. However,
when I image the air-water interface, the optical path does not include
a coverslip. I have a very limited understanding of optics, but how
might not including a coverslip affect spherical/other aberrations? Any
insight is greatly appreciated!
-Ian
Yes, you will get spherical abberation (hazyness) if the specimen is
on the surface of the water (comparable to looking at it dry). With a
layer of water on top of your sample (about 0.2mm), the correction is
about right, more then that, again spherical abberation is building
up. Depend a bit on how critically you will look at your sample, at
200x it's hard to spot.
HTH, Rene.
.
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