NetNotes
As I understand it, the issue is crystallization formed by mixing
different oils on the same microscope objective. However, any one oil used without contamination of another would be fine. Tere might be subtle optimization for the optic/aberrations that are brand-specific and might only be noticed at high resolution. I have never done a test or seen the data for crystallization, nor seen the optical differences published. Is it worth someone publishing a comparison of modern oils on different objectives? It would be great if we could all just buy inexpensive Cargille oil (which I have heard that many companies do and then rebottle) and add specific chemicals as/if needed. Ditto mounting media. Michael Abanto
michael.abanto@unibas.ch
Even relatively modern oils may form crystals on long storage. Or
some components may be slightly volatile. Ten, over years and years, you may find the refractive index is way off. Many Cargille oils mix OK. Except for the low RI ones (fluorinated hydrocarbons). Get an Abbé refractometer to measure the actual RI (measuring dispersion tends to be less precise). People are afraid of autofluorescence. Low AF is important in widefield, critical in TIRF, but confocals don’t really care. My $0.02. Zdenek Svindrych
zdedenn@gmail.com
We have posted some data on chromatic aberration on our web
site:
https://www.bioimaging.bmc.med.uni-muenchen.de/news/ chrom-ab-100x/
index.html. On our Leica SP8 STED, we checked chromatic aberration in reflection mode, relative to 470nm with the 100x1.4 STED white objective, which was manufactured for particularly low chromatic aberration. With the Zeiss 518 F oil we get up to about 50 nm, with Cargille HF up to 100 nm, and more than that with the 775 nm depletion line. STED with 775 is essentially not working with that oil, because the depletion donut is too far off in z. “Normal” NA 1.4 objectives may have an aberration of >200 nm anyway, so if you are serious about it, you will have to correct by post- processing, and then it may not matter which oil you use, because they will differ just by the amount that you have to correct for. If I have it right, the difference in behavior is described by the dispersion value, which is given as ve=something. We, however, decided to stick to the 518 F. Per slide, the additional costs are negligible, and we may save our users some trouble. And save us the trouble of users mixing different oils. Steffen Dietzel
lists@dietzellab.de
For most of our systems we use Cargille Labs type LDF. Unlike
other brands, it does not dry into a sticky coating, and provides consistency across microscopes. We can jump from scope to scope. Our users don’t mix oils. None of our users have noticed any reduction in performance and when we have checked, we were hard-pressed to measure differences. We have enough of a problem with people using Immersol W when they should be using 1.518 oil and vice versa. An exception is the Elyra, where we use Zeiss oil. Another exception is Airyscan. And, of course, the silicon lenses, but we only have one scope with a subset of users. You can try different oils; just clean completely between them. You might find real performance improvements. And if I’m wrong about Cargille LDF, please let me know! Michael Cammer
michael.cammer@
med.nyu.edu
Apart from the spherical aberration and the consequent loss
in contrast and resolution, different oils have different dispersion properties. I had quite an unexpected surprise in the past when measuring chromatic shiſt on a DELTA VISION system, first with Olympus and then with ZEISS oil. Tis aspect should not be underestimated. Davide Accardi
davide.accardi@
research.fchampalimaud.org
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Crossword Puzzle Answers See puzzle on page 74.
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