I simulated the performance of high NA objectives using both vector diffraction theory plus the Gibson and Lanni aberration model and objective lens prescriptions from the patents of various companies (Nikon, Olympus). I found that spherical aberrations are sensitive to an index variation of ~0.002 for oil and ~0.003 for the cover glass.
We had measured the dispersion of a few oils and found their Abbe numbers to be different - confirming Kurt's suspicion.
Ning
====================================
Ning Y. Chan
Nanotechnology Measurements Division
Agilent Technologies, Inc.
5301 Stevens Creek Blvd
Santa Clara, CA 95052
phone: 408 553 3123
FAX: 408 246 5925
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Kurt Thorn
Sent: Tuesday, March 17, 2009 10:11 AM
To: [log in to unmask]
Subject: Re: Immersion Oil
One thing I have heard is that different oils have different
dispersions, so you may introduce chromatic aberration if you use oils
from different vendors. I would do the experiment with multicolored
beads and look for axial shifts in the different channels before
deciding to switch oils.
Kurt
Maddox Paul wrote:
>
> It's an easy test to prove that the last (third) decimal place makes a
> reasonable difference. Using the Cargille test kit (range of ~20 RI
> around 1.515 step by 0.001), you can acquired PSFs with the different
> oils and just take a look at the x,z projections. You'll see a pretty
> dramatic shift in spherical aberration, even on small steps (ok, 0.001
> is pretty small change, but 0.002 change is measurable for sure). It
> gets even more interesting (dare I say fun) if you mount the beads in
> some low percentage agar so that you get beads at random distances
> from the coverglass! I did this one weekend and recommend it if you
> need a distraction (from grants for instance...).
>
> As for mixing oils, I always thought the big worry was that different
> brands might not be miscible and therefore you could introduce a new
> RI boundary, causing aberrations. I have never tested, or in fact seen
> this demonstrated (on purpose or otherwise). Nonetheless, I will
> continue to not mix my oils.
>
> Happy imaging,
>
> Paul
>
> Paul S. Maddox, PhD
> Assistant Professor
> Institute for Research in Immunology and Cancer
> Dept of Pathology and Cell Biol, U. de Montreal
> P.O. Box 6128, Station Centre-Ville
> Montréal QC H3C 3J7
> CANADA
>
> Courier:
> 2900, boulevard Édouard-Montpetit
> Pavillon Marcelle-Coutu, Quai 20
> Montreal QC H3T 1J4
> CANADA
>
> [log in to unmask] <mailto:[log in to unmask]>
> Ph: 514-343-7894
> Fax: 514-343-6843
>
> ------------------------------------------------------------------------
>
> *From:* Confocal Microscopy List
> [mailto:[log in to unmask]] *On Behalf Of *Craig Brideau
> *Sent:* Tuesday, March 17, 2009 12:24 PM
> *To:* [log in to unmask]
> *Subject:* Re: Immersion Oil
>
> When you are down to the third decimal place it's not going to make a
> huge difference*. You might want to try and see if you observe any
> difference between oil, but I suspect you won't see much change.
>
> Craig
>
> *unless you are talking about fiber optics or some optoelectronics...
>
> On Tue, Mar 17, 2009 at 9:58 AM, stu_the_flat <[log in to unmask]
> <mailto:[log in to unmask]>> wrote:
>
> Isn't there a 0.002 difference in the diffraction between some of the
> immersion oils?
>
> Off the top of my head
>
> Olympus Oil = 1.515
> Ziess Oil = 1.515
> Nikon Oil = 1.513
>
> I don't think it make the world of difference but as I am interested
> in PSFs
> I always match the oil the microscope.
>
> Stuart
>
>
>
>
>
> David Knecht-charter wrote:
> >
> > Being a natural skeptic, that has been my presumption as well. I
> > haven't talked to anyone who has actually seen mixed oils gum up their
> > scope or dissolve lens cement (if you believe that last one, I have a
> > few million dollars in my uncle's account in Africa I will be happy
> > to send you). Dave
> >
> > On Mar 17, 2009, at 11:37 AM, Peter Carroll wrote:
> >
> >>
> >> > I have been told by a Nikon representative that Zeiss oil could
> >> destroy the Nikon objective lens
> >>
> >> Of course you heard that... Nikon wants you to purchase /their/
> >> consumables, not their competitors! Heh...
> >
> > Dr. David Knecht
> > Department of Molecular and Cell Biology
> > Co-head Flow Cytometry and Confocal Microscopy Facility
> > U-3125
> > 91 N. Eagleville Rd.
> > University of Connecticut
> > Storrs, CT 06269
> > 860-486-2200
> > 860-486-4331 (fax)
> >
> >
> >
> >
>
> --
> View this message in context:
> http://n2.nabble.com/Immersion-Oil-tp2491973p2492239.html
> Sent from the Confocal Microscopy List mailing list archive at Nabble.com.
>
--
Kurt Thorn, PhD
Director, Nikon Imaging Center
University of California San Francisco
UCSF MC 2140
Genentech Hall Room S252
600 16th St.
San Francisco, CA 94158-2517
http://nic.ucsf.edu
phone 415.514.9709
fax 415.514.4300
|
