Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Has anyone checked the parfocality of this lens across the laser lines, say from 405 nm to 633nm, by 
reflection or multi-color beads? Asked with due regard for variability between individual lenses.

Regards,

Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923
(206) 616-4156

On Fri, 15 Jun 2007, Julio Vazquez wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>
> Eric,
>
> Regarding specifically the Zeiss alpha-Plan Fluar 100x/1.45, I find it does 
> provide the excellent resolution than one would expect from calculations, and 
> images collected with it (e.g. of fluocells stained for actin filaments or 
> mitochondria, that have very fine detail) look a bit crisper than with our 
> other other objectives, excluding perhaps the PlanApo 40x/1.2 water, which is 
> also excellent. Quantitative comparison for brightness are tricky to perform 
> accurately, but here are some of my observations:
>
> On a widefield system, when I image the same region of very thin fluorescent 
> cells (so that I can be sure to be in the very same focal plane), the Fluar 
> 100/1.45 is about 20% brighter than our older 100x/1.4 PlanApo, when I compare 
> the intensity of the same features corrected for background.
>
> On an LSM 510 system, using the same test, and keeping all things equal, 
> pinhole set to 1 for both objectives, optimal pixel size, the 100x/1.45 is also 
> about 20% brighter than the Plan Apo 100/1.4. On the other hand, our Plan 
> Neofluar 40x/1.3 is significantly brighter, since I have to increase the laser 
> power used with the 40x/1.3 by about 70-80% to get a comparable image with the 
> 100/1.45.  I suspect that part of the difference comes from the fact that the 
> 40x/1.3 has a visibly larger back aperture, and therefore may collect a greater 
> fraction of the laser beam, and therefore transmit more laser intensity to the 
> sample.
>
> In any event, if you feel the objective may not be performing properly, you may 
> want to set up your own tests. Collecting a PSF and comparing it to other 
> lenses you have (or to the theoretical expectations) is a simple and 
> informative test. The PSF should be tighter than with a 40x/1.3 for instance. 
> You also want a symmetrical PSF. If your objective is damaged, it should 
> show...
>
> --
> Julio Vazquez
> Fred Hutchinson Cancer Research Center
> Seattle, WA 98109-1024
>
>
> [log in to unmask]
> http://www.fhcrc.org/
>
>
>
>
> On Jun 15, 2007, at 12:02 AM, Michael Weber wrote:
>
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>> 
>> Dear all,
>> 
>> Mario already mentioned a couple of points. Different objectives could
>> have different pros and cons. Some of them are very bright in the green
>> range, but show poor transmission in UV and/or IR. One also needs to find
>> a good compromise between correction (spherical abberation etc.),
>> magnification, resolution and transmission for the desired wavelengths.
>> 
>> Btw Ian, compared to which objective is the 100x/1.45 Zeiss alpha-Plan
>> better? We have also one and it's less bright over the whole spectrum than
>> their 63x/1.4 Plan-Apo (which is already not the brightest, but has other
>> advantages). I'm just asking because I expected the alpha-Plan to be much
>> better, and maybe we should ask for an exchange.
>> 
>> cheers,
>> Michael
>> 
>> 
>>> Search the CONFOCAL archive at
>>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>> 
>>> 	I've just bought a Zeiss x100 alpha Plan-Fluar for my 510 meta and
>>> it's the best objective I've ever used. My knees were shaking at the sheer
>>> beauty of the images coming from the instrument. Ask for a demo and be
>>> amazed.
>>> Ian.
>>> 
>>> -----Original Message-----
>>> From: Confocal Microscopy List [mailto:[log in to unmask]] On
>>> Behalf Of Eric Scarfone
>>> Sent: 14 June 2007 16:03
>>> To: [log in to unmask]
>>> Subject: Objectives
>>> 
>>> Search the CONFOCAL archive at
>>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>> 
>>> Hello all
>>> Please forgive me if these kind of issues are not discussed here.
>>> While testing for new microscopes I had the strong impression that high
>>> mag optics are much more luminous and crisp with Nikon than with Zeiss
>>> (ex: Nikon 60x/1.4 versus Zeiss 63x/1,4 plan apo).
>>> I had the exact oposite impression using their plan apo 10x/0,45
>>> 
>>> Of course this is very subjective and I'd like to know if some of you
>>> are aware of more objective differences (i.e. in the making of these
>>> objectives) that would sustain/negate my impressions.
>>> Best Regards to all and congratulations for keeping this list so active
>>> Eric
>>> 
>>> 
>>> Eric Scarfone, PhD, CNRS,
>>> Center for Hearing and communication Research
>>> Department of Clinical Neuroscience
>>> Karolinska Institutet
>>> 
>>> Postal Address:
>>> CFH, M1:02
>>> Karolinska Hospital,
>>> SE-171 76 Stockholm, Sweden
>>> 
>>> Work:  +46 (0)8-517 70343,
>>> Fax:   +46 (0)8-301876
>>> 
>>> 
>>> http://www.ki.se/cfh/
>>> 
>
>