Re: Zeiss 40X N.A. 1.4 Plan APo as replacement for 63X?

Posted by Miroslav Varecha on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Zeiss-40X-N-A-1-4-Plan-APo-as-replacement-for-63X-tp7582324p7582335.html

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Hello,
I would like to react to George. In my humble opinion, best lateral
optical resolution for confocal microscope is around 130 nm, so going
to resolution 60x60 nm is overshoot and it is just wasting your drive
space as you are not collecting any new real information. Resolutions
of 50x50nm and such are area of superresolution microscopy. Our
confocal Zeiss LSM 700 we have also highest resolution of 2k x 2k.
High quality CCDs have even less, but sCMOS can get higher than 2k x
2k for sure. We have objective Zeiss 40x but NA 1,3 and I can tell you
that it is the most used objective (usually we observe stem cells)
even tho it is not NA1,4. We have also 63x1,4 NA but ppl prefer larger
field of view of 40x. You still get great detail and many cells in one
image. But if you want to observe really up close organels and such I
would go to 63x NA1,4. 40x NA 1,4 seems to me like most flexible
objective for vast majority of observations.
Miroslav


2014-07-14 1:31 GMT+02:00 George McNamara <[hidden email]>:

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> Hi Michael,
> why 2kx2k?
> If the 40x lens has a 250x250 um field of view, this would be undersampling,
> pixel size 125x125 nm. If even larger field of view, undersampling even
> more.
> I suggest pixel size of 50x50 or 60x60 nm, and 3D deconvolution (Z step 200
> nm, maybe closer),
> George
>
> On 7/11/2014 8:53 AM, Cammer, Michael wrote:
>>
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>>
>> Does anyone have experience with the new Zeiss 40X N.A. 1.4 PlanApo?  This
>> is something I've wanted for a long time, the ability to take large fields
>> of view (2k X 2k pixels) at high resolution instead of having to do tiling.
>> Also, with the new cameras that have oodles of small pixels...
>>
>> I'm considering replacing our 63X with this new 40X.  Any experience with
>> this, other than the battle of having to explain to other scope users why
>> this is not really lower magnification?
>>
>> Regards,
>>
>> Michael
>>
>>
>> ===========================================================================
>> Michael Cammer, Microscopy Core&  Dustin Lab , Skirball Institute, NYU
>> Langone Medical Center
>> Cell:  914-309-3270   Lab: 212-263-3208
>> http://ocs.med.nyu.edu/microscopy&
>> http://www.med.nyu.edu/skirball-lab/dustinlab/
>>
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> George McNamara, Ph.D.
> Single Cells Analyst
> L.J.N. Cooper Lab
> University of Texas M.D. Anderson Cancer Center
> Houston, TX 77054
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