Posted by
George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Experience-with-Olympus-X-Line-100X-objective-tp7592261p7592262.html
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Hi Abby,
I demo'd X-Line when it was just introduced - though only for a brief
visit here at JHU (
http://confocal.jhu.edu/current-equipment/fishscope ).
Yes, excellent lenses.
Frankly, any brand new objective lens will work better than any you've
used extensively.
To get the benefit of 1.45 NA in the absence of through objective lens
TIRF excitation, you need to optimize the N.A. of the specimen, i.e.
R.I. 1.518. Possibly also 170 um coverglass thickness (though in theory,
if R.I. of glass, immersion medium, specimen medium is the same, could
use thinner coverglass and image deeper ... of course the specimen
itself should be perfect R.I. match too, i.e. Expansion Microscopy).
Big unknown: Price difference between a standard 100x (1.40 or 1.45) and
the X-Line. If X is 2X price, for a few percent gain (with perfect
specimen), then probably not going to be worth it.
I suggest: discuss with your Olympus sales rep getting quote and demo of
'standard' and the X-Line.
*
Potentially useful comparison:
https://svi.nl/NyquistCalculatorfor perfect lens, exactly R.I. 1.515 (or 1.518) for immersion and
specimen R.I.s, ex 488nm, em 520nm, widefield (implicitly intending to
do deconvolution)
1.45 NA perfect lens:
x:89nm (this or smaller pixel size)
y:89nm (this or smaller Z-step size).
z:241nm
Use the web page to see XY and XZ images of PSF.
1.40 NA perfect lens:
x:92nm
y:92nm
z:277nm
So, the +0.05 NA gets nominally 3 nm in XY, 36 nm in Z. For comparison,
one AausFP1 GFP molecule is 4.2 nm tall (addgene expression plasmd could
be a good investment too).
***
Hmmm ... if you have money, consider improving your computer. For
example, if a Windows 10 PC, NVMe M.2 SSD PCIe card (ASUS or ASROCK
Hyper M2 PCIe, whatever 1TB each NVMe drives) are typically ~3000 MB/sec
(PCIe gen3, ... gen4 can be 2x faster) vs single SATA-6 SSD ~600 MB/sec
or single HDD 100 - 200 MB/sec (note: ASUS and ASROCK require
motherboard with BIOS set for the PCIe slot to be x4x4x4x4 bifurcation
... GloTrends has an 4-Bay M.2 NVME Adapter, that does not use
bifurcation ... amazon currently lists as unavailable). More RAM in PC
good ... new GPU good (if Deltavision can use it, and if you can find an
RTX 30x0). A new PCIe gen4 PC could also be nice, and could buy as a
bundle to get an RTX 30x0 with it.
Happy 2021,
George
On 5/20/2021 3:20 PM, Abby Dernburg wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
>
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy> Post images on
http://www.imgur.com and include the link in your posting.
> *****
>
> Hi all -
>
> I am thinking of adding an Olympus X-Line 100X NA 1.45 to our DeltaVision Elite system and am curious about real-world experience with this objective, particularly for wide-field deconvolution microscopy. I haven't found any descirptions of performance other than marketing materials so I'm wondering how it measures up. Thanks in advance for any feedback!
>
> -Abby