Dear Dr. Hertzler:
Ask Wally Clark.
Steven Wang
 
 
 
On Feb 22,  3:27pm, Phil Hertzler wrote:
> Subject: Re: lenses
> BROWNVM.brown.edu and David Berk wrote:
>
> > A researcher in our department wants to use a motorized xy stage on a
confocal
> >  microscope (in combination with an xz motor) to collect data for 3D
> > reconstructions.
> > This would involve collecting four areas per embryo and making a montage
> > to get the entire embryo. One of her concerns is finding a high quality
> > low power objective (to avoid any distortion). Has anyone evaluated 10X
> > or 20X plan-apo objectives from different manufacturers?
> >
> >
> >>  I don't have any information about low power objectives, but I'm
interested
> >>in seeing the response to this question because I'd also like to do optical
> >>sectioning at low magnification.  From my understanding of optics and my
> >>limited experience with confocal microscopes I thought that with low NA
> >>objectives there is very little advantage to confocal illumination.  Has
> >>anyone documented the actual improvements in axial or lateral resolution
with
> >>low NA objectives?
>
> I used an Olympus 20X 0.7 NA (DPlanApo20UV) for confocal sections and 3D
> reconstructions of embryos approximately 150 um in diameter.  I could get
> all the way through the embryo with this dry objective, optical sectioning
> was great, and the resolution of antibody-stained microtubules was very
> good (check _Development_ 116:127-140 to make your own evaluation).  I
> haven't compared other lenses, but I sure miss that lens in my new lab!
>
> Phil Hertzler
>
>
> Duke University
> Developmental, Cell &
>   Molecular Biology Group
> B362 LSRC Building
> Box 91000
> Durham, NC  27708-1000
>
> [log in to unmask]
>-- End of excerpt from Phil Hertzler