Recommendation for 40X NA 1.4 oil lens

[Valerie Lawrence]

Hello all,

 

We have a Zeiss LSM 510 confocal microscope (inverted) system and our primary imaging lens is a Plan-Apo 63X/1.4 oil lens.  We are contemplating the purchase of a Plan-Apo 40x/1.4 oil lens for the wider field of view and ask for opinions on such a purchase.  There are two Plan-Apo 40X oil lenses we’re considering; 40X/1.4 and 40X/1.3 (which has a longer working distance but is more expensive).  We’d appreciate feedback from other users as to their recommendations and experiences.

 

Many thanks.

 

Valerie Lawrence

FPInnovations - Paprican Division

3800 Wesbrook Mall

Vancouver  BC  V6S 2L9

( : 604.222.3201 #312 | 7 : 604.222.3207 | *:[hidden email]

 

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[Martin Wessendorf-2]

Val Lawrence wrote:

> We have a Zeiss LSM 510 confocal microscope (inverted) system and our
> primary imaging lens is a Plan-Apo 63X/1.4 oil lens.  We are
> contemplating the purchase of a Plan-Apo 40x/1.4 oil lens for the wider
> field of view and ask for opinions on such a purchase.  There are two
> Plan-Apo 40X oil lenses we’re considering; 40X/1.4 and 40X/1.3 (which
> has a longer working distance but is more expensive).  We’d appreciate
> feedback from other users as to their recommendations and experiences.

Several thoughts:

1)  All else being equal, go with the 1.4 NA.  However, if you're using
an inverted stand because users are sometimes looking at live cells,
wouldn't a water-immersion lens be a better choice?  If for other
reasons you want oil but you're sometimes looking at live cells, I'd go
with the 1.3 NA since you'll end up with a bit less spherical
aberration--there will be fewer rays entering from extreme angles.

2)  If you have users who look at thick preparations (e.g. insect whole
mounts, filled cells from slice electrophysiology experiments,
inexperienced histologists who slop on too much mounting medium, etc.),
  the greater working distance may be an important advantage.

3)  Never trust the specifications for working distance--measure it
yourself.  The specified values may not necessarily be accurate.  You
should be able to get a rough idea of working distance by touching the
objective to the surface of a mirror slide (--look for the barrel of the
objective beginning to telescope in on itself--don't over-do it and
break the slide!) and then defocussing till you get a reflectance image.
   A more precise method would be to mount specimens with spacers of
known thickness under the coverglass--that allows you to compare the two
objectives directly.

Good luck!

Martin
--
Martin Wessendorf, Ph.D.                   office: [phone]
Assoc Prof, Dept Neuroscience                 lab: [phone]
University of Minnesota             Preferred FAX: [phone]
6-145 Jackson Hall, 321 Church St. SE    Dept Fax: [phone]
Minneapolis, MN  55455                    e-mail: [hidden email]

[Julio Vazquez]

Hi Valerie, 

If you have an older Zeiss 510, chances are you are using the older Zeiss  objective mount (W 0.8). I think the PlanApo 40/1.4 is a newer objective in the new Zeiss M27 mount, so this may not be an option for you. Zeiss has adapters to mount older objectives on the newer scopes, but I don't know if they have the reverse adapter..., the new M27  objectives have a larger back aperture, so you would probably be losing some performance by using those on your scope. Therefore, the older 40/1.3 is probably your best choice (and you get a longer working distance, which is typically a bonus). Maybe they have a PlanApo 40x/1.4 in the older mount which I am not aware of, but even so, I would think that the extra working distance of the 1.3 might be more useful than the marginal increase in resolution and brightness of the 1.4, but I really don't know since I haven't compared them. On a newer scope, the new lens is probably better, both because of the higher NA, and the newer design. This is the case for the new 20x/0.8 which is visibly better than the older 20x/0.75.

Incidentally, the Zeiss Plan Neofluar 40/1.3 oil is also a pretty good lens for confocal, in my opinion. It's probably cheaper than the PlanApo, and chances are it might be brighter too, although I do not have a PlanApo to compare. Chromatic correction for Plan Neofluar is pretty good and we do use it a lot on our LSM 510. 

The Zeiss web site has good info about their objectives (transmission curves, color correction, etc, ), so you may want to have a look and decide which one is best based on your application, the wavelengths you use, etc....

http://www.zeiss.de/C12567BE0045ACF1/Contents-Frame/A22821A1BDCAAE49C12574E80030638E

--

Julio Vazquez

Fred Hutchinson Cancer Research Center

1100 Fairview Ave. N.,  mailstop DE-512

Seattle, WA 98109-1024

http://www.fhcrc.org/

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On May 14, 2010, at 11:49 AM, Val Lawrence wrote:

Hello all,

 

We have a Zeiss LSM 510 confocal microscope (inverted) system and our primary imaging lens is a Plan-Apo 63X/1.4 oil lens.  We are contemplating the purchase of a Plan-Apo 40x/1.4 oil lens for the wider field of view and ask for opinions on such a purchase.  There are two Plan-Apo 40X oil lenses we’re considering; 40X/1.4 and 40X/1.3 (which has a longer working distance but is more expensive).  We’d appreciate feedback from other users as to their recommendations and experiences.

 

Many thanks.

 

Valerie Lawrence

[George McNamara]

Hi Valerie,

No reason to make a blind purchase - test both, on your microscope.

Possibly wait for the Jim Pawley course to arrive in town, along with a bunch of Zeiss reps and lots of lenses.

as for working distance, start using 170 um coverglasses instead of generic #1.5 coverglasses (the latter's range 160 to 190 um). If you use perfectly 1.51x refractive index matched mounting media - such as 2,2'-thiodiethanol (Staudt ... Hell paper), in principle could switch to #1 or #0 coverglasses to reach deeper into the specimen (fixed cells).

George


At 02:49 PM 5/14/2010, you wrote:

Hello all,
 
We have a Zeiss LSM 510 confocal microscope (inverted) system and our primary imaging lens is a Plan-Apo 63X/1.4 oil lens.  We are contemplating the purchase of a Plan-Apo 40x/1.4 oil lens for the wider field of view and ask for opinions on such a purchase.  There are two Plan-Apo 40X oil lenses we’re considering; 40X/1.4 and 40X/1.3 (which has a longer working distance but is more expensive).  We’d appreciate feedback from other users as to their recommendations and experiences.
 
Many thanks.
 
Valerie Lawrence
FPInnovations - Paprican Division
3800 Wesbrook Mall
Vancouver  BC  V6S 2L9
( : 604.222.3201 #312 | 7 : 604.222.3207 | * : [hidden email]
 

George McNamara, Ph.D.
Image Core Manager
Analytical Imaging Core Facility
University of Miami, Miller School of Medicine
Miami, FL 33136
[hidden email]
[hidden email]
305-243-8436 office
http://www.sylvester.org/AICF (Analytical Imaging Core Facility)
http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file)
http://home.earthlink.net/~geomcnamara