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
Courier:
2900, boulevard Édouard-Montpetit
Pavillon Marcelle-Coutu, Quai 20
[hidden email]
Ph: 514-343-7894
Fax: 514-343-6843
From:
Sent: Tuesday, March 17, 2009
12:24 PM
To:
[hidden email]
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 <[hidden email]> wrote:
Isn't there a 0.002 difference in the diffraction between some of the
immersion oils?
Off the top of my head
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
> 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
>
>
>
> 860-486-2200
> 860-486-4331 (fax)
>
>
>
>
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