Posted by
James Pawley on
URL: http://confocal-microscopy-list.275.s1.nabble.com/resolution-of-Zeiss-Water-and-Glycerine-63X-lens-tp7099174p7101356.html
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>Hi, everyone:
>
>I'm looking for a 63X objective for live imaging and its resolution
>is my major concern. Zeiss recommands Objective LCI Plan-Neofluar
>63x/1.3 Imm Corr DIC M27, saying it has NA = 1.3 in both water and
>glycerine. Does it really have NA=1.3 even in water?
>
>Your reply is highly appreciated.
>Xiao
>
>*******************
>Xiao Liu
>School of Life Sciences
>Tsing-Hua University
>Beijing, China
>*******************
>
Well, it may have such an NA in water, but most of the rays
approaching the flat surfaces (two sides of the cover slip and the
front element of the objective) will be reflected when transmitting
these surface, so will not contribute much to the image.
Keep in mind that, when actually tested, using the Methods in Chapter
11 of the Handbook, Juskaitis found that even high quality NA 1.4
objectives only worked "properly" up to 1.35 (The high-NA ray were
indeed propagated by the lenses but had the wrong phase so actually
did not contribute to image sharpness).
In general, when viewing living specimens, "resolution" (that
slippery concept that can only be considered in terms of more or less
contrast) is more likely degraded by aberrations brought on by wrong
immersion media or by the RI variations of the specimen itself than
by the NA marked on the side of the lens.
Cheers,
Jim Pawley
--
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