Experience with Olympus X-Line 100X objective?

[Abby Dernburg]

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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

[George McNamara]

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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/NyquistCalculator

for 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

[Ferhan A]

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Hi Abby,



For our spinning disk microscope, we've recently purchased the exact
Olympus X-line model you mentioned:

UPLXAPO100XO (N.A 1.45, FN 26.5, coverslip 0.17, WD 0.13mm)

Earlier we were using:

UPLFLN100XO (N.A: 1.3, FN26.5, coverslip 0.17, WD 0.2mm)

(We put the old one back to Olympus Cell-R microscope from which we
borrowed it after the arrival of the new spinning disk microscope)

The old one has a slightly longer working distance (0.2 vs 0.13mm), but the
new X-line objective is clearly brighter and has better resolution (NA 1.45
vs 1.3). In our case, it significantly improved the performance of our
spinning disk microscope.

Regards,

Ferhan


---
Ferhan Ayaydin, Ph.D.
---

*Head of Advanced Core Facility *HCEMM, Hungarian Centre of Excellence for
Molecular Medicine
Functional Cell Biology and Immunology Advanced Core Facility
University of Szeged, Faculty of Medicine, 6720 Szeged, Hungary
---

* Senior research associate *Biological Research Centre, Cellular Imaging
Laboratory
Temesvári krt. 62, 6726 Szeged, Hungary
---

* Research associate *University of Szeged, Faculty of Medicine,
Albert Szent-Györgyi Health Centre
Interdisciplinary R&D and Innovation Center of Excellence
Department of Dermatology and Allergology,
Korányi fasor 6, 6720 Szeged, Hungary



On Thu, May 20, 2021 at 9:30 PM Abby Dernburg <[hidden email]>
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
>

[Alison J. North-2]

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Hi Abby,

I don't know whether this will be useful information or not, but we recently purchased the 60X X-line objective for our DeltaVision, and I can confirm that the psfs look great and the chromatic aberrations are significantly better than with the original 60x/1.42 oil objective, even when using the Cargille oils (which result in noticeable chromatic dispersion in comparison to when using Olympus oil).  If the 100x X-line objective is anything like as good, then I really should be thinking about buying one myself - with the new 60x we no longer need to do any post-acquisition corrections.

All the best,
Alison

Alison J. North, Ph.D.
Senior Director of the Bio-Imaging Resource Center and Research Associate Professor
The Rockefeller University
1230 York Avenue
New York
NY 10065
Tel. [phone] (office - direct line)
https://www.rockefeller.edu/bioimaging/
RRID:SCR_017791

________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Abby Dernburg <[hidden email]>
Sent: Thursday, May 20, 2021 3:20 PM
To: [hidden email] <[hidden email]>
Subject: Experience with Olympus X-Line 100X objective?

*****
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Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=NmNoIJspXtyELNquCYzD3rsekE-Q2HTeMGZalX8tkAE&s=8bFDOVYQ_MjyauSQQ8n-kZzEw04Py7NnwYipjGLnjM0&e=  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

[Owens, Peter]

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I have just purchased a 100x  xline for our facility in NUIG , to be delivered in a few weeks . Interesting to read your comments Alison on the psf and chromatic aberration. Certainly looking forward to trying it out and I can report back to this forum when i have a chance to use it.  I have also heard that Olympus pricing for optics is due to increase by around 20% in June , and this was already a pricey objective.

Best regards

Peter

www.imaging.nuigalway.ie<http://www.imaging.nuigalway.ie>
Centre for Microscopy and Imaging at NUI Galway - current research<http://www.imaging.nuigalway.ie/>
The Centre for Microscopy and Imaging at NUIG (CMI) is a core facility dedicated to providing scientists with a variety of high-end light and electron microscopes along cutting-edge imaging software and analysis tools. Provide expertise in microscopy, imaging and analysis specialising in biomaterials, cell and tissue phenotypes.
www.imaging.nuigalway.ie




________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Alison J. North <[hidden email]>
Sent: 21 May 2021 11:03
To: [hidden email] <[hidden email]>
Subject: Re: Experience with Olympus X-Line 100X objective?

*****
To join, leave or search the confocal microscopy listserv, go to:
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Hi Abby,

I don't know whether this will be useful information or not, but we recently purchased the 60X X-line objective for our DeltaVision, and I can confirm that the psfs look great and the chromatic aberrations are significantly better than with the original 60x/1.42 oil objective, even when using the Cargille oils (which result in noticeable chromatic dispersion in comparison to when using Olympus oil).  If the 100x X-line objective is anything like as good, then I really should be thinking about buying one myself - with the new 60x we no longer need to do any post-acquisition corrections.

All the best,
Alison

Alison J. North, Ph.D.
Senior Director of the Bio-Imaging Resource Center and Research Associate Professor
The Rockefeller University
1230 York Avenue
New York
NY 10065
Tel. [phone] (office - direct line)
https://www.rockefeller.edu/bioimaging/
RRID:SCR_017791

________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Abby Dernburg <[hidden email]>
Sent: Thursday, May 20, 2021 3:20 PM
To: [hidden email] <[hidden email]>
Subject: Experience with Olympus X-Line 100X objective?

*****
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Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=NmNoIJspXtyELNquCYzD3rsekE-Q2HTeMGZalX8tkAE&s=8bFDOVYQ_MjyauSQQ8n-kZzEw04Py7NnwYipjGLnjM0&e=  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

[Abby Dernburg]

*****
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*****

Thanks to those who responded - now I’m excited to try out the X-line (and thinking I should consider the 60X as well). I didn’t realize that Olympus oil might have been a better choice than the Cargille oils we’ve been using - thanks for that tip, Alison. We do see pretty severe chromatic dispersion in Z with our current 100X objective.

Abby Dernburg, Ph.D.
HHMI Investigator
Department of Molecular and Cell Biology
University of California, Berkeley
Berkeley, CA 94720
USA

Mobile: [phone]

> On May 21, 2021, at 3:30 AM, Owens, Peter <[hidden email]> 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.
> *****
>
> I have just purchased a 100x  xline for our facility in NUIG , to be delivered in a few weeks . Interesting to read your comments Alison on the psf and chromatic aberration. Certainly looking forward to trying it out and I can report back to this forum when i have a chance to use it.  I have also heard that Olympus pricing for optics is due to increase by around 20% in June , and this was already a pricey objective.
>
> Best regards
>
> Peter
>
> www.imaging.nuigalway.ie<http://www.imaging.nuigalway.ie>
> Centre for Microscopy and Imaging at NUI Galway - current research<http://www.imaging.nuigalway.ie/>
> The Centre for Microscopy and Imaging at NUIG (CMI) is a core facility dedicated to providing scientists with a variety of high-end light and electron microscopes along cutting-edge imaging software and analysis tools. Provide expertise in microscopy, imaging and analysis specialising in biomaterials, cell and tissue phenotypes.
> www.imaging.nuigalway.ie
>
>
>
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on behalf of Alison J. North <[hidden email]>
> Sent: 21 May 2021 11:03
> To: [hidden email] <[hidden email]>
> Subject: Re: Experience with Olympus X-Line 100X objective?
>
> *****
> 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 Abby,
>
> I don't know whether this will be useful information or not, but we recently purchased the 60X X-line objective for our DeltaVision, and I can confirm that the psfs look great and the chromatic aberrations are significantly better than with the original 60x/1.42 oil objective, even when using the Cargille oils (which result in noticeable chromatic dispersion in comparison to when using Olympus oil).  If the 100x X-line objective is anything like as good, then I really should be thinking about buying one myself - with the new 60x we no longer need to do any post-acquisition corrections.
>
> All the best,
> Alison
>
> Alison J. North, Ph.D.
> Senior Director of the Bio-Imaging Resource Center and Research Associate Professor
> The Rockefeller University
> 1230 York Avenue
> New York
> NY 10065
> Tel. [phone] (office - direct line)
> https://www.rockefeller.edu/bioimaging/
> RRID:SCR_017791
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on behalf of Abby Dernburg <[hidden email]>
> Sent: Thursday, May 20, 2021 3:20 PM
> To: [hidden email] <[hidden email]>
> Subject: Experience with Olympus X-Line 100X objective?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=NmNoIJspXtyELNquCYzD3rsekE-Q2HTeMGZalX8tkAE&s=PXZ7op5XSrDNBV28-AjO-FAY-PMyeGqCS8w3UOpmlAQ&e=
> Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=NmNoIJspXtyELNquCYzD3rsekE-Q2HTeMGZalX8tkAE&s=8bFDOVYQ_MjyauSQQ8n-kZzEw04Py7NnwYipjGLnjM0&e=  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

[Alison J. North-2]

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Hi Abby,

So the situation with the Olympus objectives and different oils has been an interesting journey for me.  And I'm happy to have an opportunity to publicly set the records straight because I previously commented that the Olympus 100x SApo - which is described as being corrected across all regular wavelength dyes - appeared to give chromatic shifts. Like you, I use DeltaVisions and an OMX a lot.  (Of course, you don't really notice the chromatic aberrations with the OMX because you use the image registration step to correct for the multiple cameras anyway.)  With the DeltaVisions we always had to correct for chromatic shift when imaging at high resolution, e.g. for yeast or bacteria, and I blamed this entirely on the objectives.  It never occurred to me that the Cargille oils, which we always use to compensate for refractive index mismatch, could be the issue, until somebody at GE told me a couple of years ago that it seemed that the composition of the Cargille oils had changed over the past few years and now they appeared to induce chromatic dispersion.  Therefore, when I tested the new X-line 60x objective against the old 60x, I did so both using Cargille oil and standard Olympus oil, and that's how I confirmed that indeed the shift was far more pronounced with the Cargille oil for the old objective.

Of course, this leaves one with a bit of a dilemma - I would still want to use the Cargille oils because of their fabulous range in r.i.  that allows you to minimize spherical aberrations.  So I was delighted to find that with the X-line 60x I can stick to the Cargille oils without worrying about chromatic aberrations.  And I owe a public apology to Olympus for having maligned their SApo objectives unfairly!

I do think it's an important lesson.  People are all too happy to switch oils around without having thoroughly tested the consequences.

I'd be interested to hear how you get on with the 100x/1.45 X-line objective - I'm certainly sold on the 40x and 60x versions!

All the best,
Alison




________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Abby Dernburg PhD <[hidden email]>
Sent: Friday, May 21, 2021 11:02 AM
To: [hidden email] <[hidden email]>
Subject: Re: Experience with Olympus X-Line 100X objective?

*****
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Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=HW0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=oViXu4_HYXZj9AXrvYXRTNWfO1-HND9lhMlpp03OmzE&e=  and include the link in your posting.
*****

Thanks to those who responded - now I’m excited to try out the X-line (and thinking I should consider the 60X as well). I didn’t realize that Olympus oil might have been a better choice than the Cargille oils we’ve been using - thanks for that tip, Alison. We do see pretty severe chromatic dispersion in Z with our current 100X objective.

Abby Dernburg, Ph.D.
HHMI Investigator
Department of Molecular and Cell Biology
University of California, Berkeley
Berkeley, CA 94720
USA

Mobile: [phone]

> On May 21, 2021, at 3:30 AM, Owens, Peter <[hidden email]> wrote:
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=HW0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=QNZ1Vxc7NHyZ7J-pr29txEQeUdZo1Q8N09PPFl4KVEg&e=
> Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=HW0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=oViXu4_HYXZj9AXrvYXRTNWfO1-HND9lhMlpp03OmzE&e=  and include the link in your posting.
> *****
>
> I have just purchased a 100x  xline for our facility in NUIG , to be delivered in a few weeks . Interesting to read your comments Alison on the psf and chromatic aberration. Certainly looking forward to trying it out and I can report back to this forum when i have a chance to use it.  I have also heard that Olympus pricing for optics is due to increase by around 20% in June , and this was already a pricey objective.
>
> Best regards
>
> Peter
>
> www.imaging.nuigalway.ie<https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imaging.nuigalway.ie&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=HW0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=98RawYRKoHtC_g9VjPzmyX23HHHO_Ppk2sf1aEY0dNE&e=<http://www.imaging.nuigalway.ie<https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imaging.nuigalway.ie&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=HW0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=98RawYRKoHtC_g9VjPzmyX23HHHO_Ppk2sf1aEY0dNE&e=> >
> Centre for Microscopy and Imaging at NUI Galway - current research<https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imaging.nuigalway.ie_&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=HW0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=2eyZOxnAR68xtkfRupUZaUnuhMXgQ7RLJYoQ8xGvpxQ&e= >
> The Centre for Microscopy and Imaging at NUIG (CMI) is a core facility dedicated to providing scientists with a variety of high-end light and electron microscopes along cutting-edge imaging software and analysis tools. Provide expertise in microscopy, imaging and analysis specialising in biomaterials, cell and tissue phenotypes.
> www.imaging.nuigalway.ie<http://www.imaging.nuigalway.ie>
>
>
>
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on behalf of Alison J. North <[hidden email]>
> Sent: 21 May 2021 11:03
> To: [hidden email] <[hidden email]>
> Subject: Re: Experience with Olympus X-Line 100X objective?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=HW0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=oViXu4_HYXZj9AXrvYXRTNWfO1-HND9lhMlpp03OmzE&e=  and include the link in your posting.
> *****
>
> Hi Abby,
>
> I don't know whether this will be useful information or not, but we recently purchased the 60X X-line objective for our DeltaVision, and I can confirm that the psfs look great and the chromatic aberrations are significantly better than with the original 60x/1.42 oil objective, even when using the Cargille oils (which result in noticeable chromatic dispersion in comparison to when using Olympus oil).  If the 100x X-line objective is anything like as good, then I really should be thinking about buying one myself - with the new 60x we no longer need to do any post-acquisition corrections.
>
> All the best,
> Alison
>
> Alison J. North, Ph.D.
> Senior Director of the Bio-Imaging Resource Center and Research Associate Professor
> The Rockefeller University
> 1230 York Avenue
> New York
> NY 10065
> Tel. [phone] (office - direct line)
> https://www.rockefeller.edu/bioimaging/
> RRID:SCR_017791
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on behalf of Abby Dernburg <[hidden email]>
> Sent: Thursday, May 20, 2021 3:20 PM
> To: [hidden email] <[hidden email]>
> Subject: Experience with Olympus X-Line 100X objective?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=NmNoIJspXtyELNquCYzD3rsekE-Q2HTeMGZalX8tkAE&s=PXZ7op5XSrDNBV28-AjO-FAY-PMyeGqCS8w3UOpmlAQ&e=
> Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=NmNoIJspXtyELNquCYzD3rsekE-Q2HTeMGZalX8tkAE&s=8bFDOVYQ_MjyauSQQ8n-kZzEw04Py7NnwYipjGLnjM0&e=  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

[VERMEREN Matthieu]

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Hi Alison,

It's funny you should say that because I've just learned it here:

https://www.microscope.healthcare.nikon.com/en_EU/learn/video-tutorials

Choose "Objective lenses and Optical Aberrations" and go to 15.04.

According to Nikon, it's not just refractive index that matters, but also dispersion that must be matched to the lens glass...

Sincerely,

Matthieu

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Alison J. North
Sent: 21 May 2021 19:04
To: [hidden email]
Subject: Re: Experience with Olympus X-Line 100X objective?

This email was sent to you by someone outside the University.
You should only click on links or attachments if you are certain that the email is genuine and the content is safe.

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Hi Abby,

So the situation with the Olympus objectives and different oils has been an interesting journey for me.  And I'm happy to have an opportunity to publicly set the records straight because I previously commented that the Olympus 100x SApo - which is described as being corrected across all regular wavelength dyes - appeared to give chromatic shifts. Like you, I use DeltaVisions and an OMX a lot.  (Of course, you don't really notice the chromatic aberrations with the OMX because you use the image registration step to correct for the multiple cameras anyway.)  With the DeltaVisions we always had to correct for chromatic shift when imaging at high resolution, e.g. for yeast or bacteria, and I blamed this entirely on the objectives.  It never occurred to me that the Cargille oils, which we always use to compensate for refractive index mismatch, could be the issue, until somebody at GE told me a couple of years ago that it seemed that the composition of the Cargille oils had changed over the past few years and now they appeared to induce chromatic dispersion.  Therefore, when I tested the new X-line 60x objective against the old 60x, I did so both using Cargille oil and standard Olympus oil, and that's how I confirmed that indeed the shift was far more pronounced with the Cargille oil for the old objective.

Of course, this leaves one with a bit of a dilemma - I would still want to use the Cargille oils because of their fabulous range in r.i.  that allows you to minimize spherical aberrations.  So I was delighted to find that with the X-line 60x I can stick to the Cargille oils without worrying about chromatic aberrations.  And I owe a public apology to Olympus for having maligned their SApo objectives unfairly!

I do think it's an important lesson.  People are all too happy to switch oils around without having thoroughly tested the consequences.

I'd be interested to hear how you get on with the 100x/1.45 X-line objective - I'm certainly sold on the 40x and 60x versions!

All the best,
Alison




________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Abby Dernburg PhD <[hidden email]>
Sent: Friday, May 21, 2021 11:02 AM
To: [hidden email] <[hidden email]>
Subject: Re: Experience with Olympus X-Line 100X objective?

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Post images on https://urldefense.proofpoint.com/v2/url?u=http-3A__www.imgur.com&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYibK1YmxXez1G3oNStg&r=RBx0-WJrAO5vwSOLNmFbqYvikvIZS5ns3-USwvMOuLo&m=HW0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=oViXu4_HYXZj9AXrvYXRTNWfO1-HND9lhMlpp03OmzE&e=  and include the link in your posting.
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Thanks to those who responded - now I’m excited to try out the X-line (and thinking I should consider the 60X as well). I didn’t realize that Olympus oil might have been a better choice than the Cargille oils we’ve been using - thanks for that tip, Alison. We do see pretty severe chromatic dispersion in Z with our current 100X objective.

Abby Dernburg, Ph.D.
HHMI Investigator
Department of Molecular and Cell Biology University of California, Berkeley Berkeley, CA 94720 USA

Mobile: [phone]

> On May 21, 2021, at 3:30 AM, Owens, Peter <[hidden email]> wrote:
>
> *****
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> *****
>
> I have just purchased a 100x  xline for our facility in NUIG , to be delivered in a few weeks . Interesting to read your comments Alison on the psf and chromatic aberration. Certainly looking forward to trying it out and I can report back to this forum when i have a chance to use it.  I have also heard that Olympus pricing for optics is due to increase by around 20% in June , and this was already a pricey objective.
>
> Best regards
>
> Peter
>
> www.imaging.nuigalway.ie<https://urldefense.proofpoint.com/v2/url?u=ht
> tp-3A__www.imaging.nuigalway.ie&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoWYib
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> www.imaging.nuigalway.ie<http://www.imaging.nuigalway.ie>
>
>
>
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Alison J. North <[hidden email]>
> Sent: 21 May 2021 11:03
> To: [hidden email]
> <[hidden email]>
> Subject: Re: Experience with Olympus X-Line 100X objective?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
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> 0j46tfOJaTyP8RjuJ5ZP_MfnqTN-q-huPyoNULrE0&s=QNZ1Vxc7NHyZ7J-pr29txEQeUd
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> *****
>
> Hi Abby,
>
> I don't know whether this will be useful information or not, but we recently purchased the 60X X-line objective for our DeltaVision, and I can confirm that the psfs look great and the chromatic aberrations are significantly better than with the original 60x/1.42 oil objective, even when using the Cargille oils (which result in noticeable chromatic dispersion in comparison to when using Olympus oil).  If the 100x X-line objective is anything like as good, then I really should be thinking about buying one myself - with the new 60x we no longer need to do any post-acquisition corrections.
>
> All the best,
> Alison
>
> Alison J. North, Ph.D.
> Senior Director of the Bio-Imaging Resource Center and Research
> Associate Professor The Rockefeller University
> 1230 York Avenue
> New York
> NY 10065
> Tel. [phone] (office - direct line)
> https://www.rockefeller.edu/bioimaging/
> RRID:SCR_017791
>
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Abby Dernburg <[hidden email]>
> Sent: Thursday, May 20, 2021 3:20 PM
> To: [hidden email]
> <[hidden email]>
> Subject: Experience with Olympus X-Line 100X objective?
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> https://urldefense.proofpoint.com/v2/url?u=http-3A__lists.umn.edu_cgi-
> 2Dbin_wa-3FA0-3Dconfocalmicroscopy&d=DwIFaQ&c=JeTkUgVztGMmhKYjxsy2rfoW
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> *****
>
> 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

The University of Edinburgh is a charitable body, registered in Scotland, with registration number SC005336. Is e buidheann carthannais a th’ ann an Oilthigh Dhùn Èideann, clàraichte an Alba, àireamh clàraidh SC005336.

[Stanislav Vitha-2]

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It is good to hear about the 60x X-line objective, but I would say that the old PLAPON 60x/1.42 does not set the bar very high. Perhaps it was better suited for wide-field observation., but when we first used it on our Fluoview system (2006), even our new users noticed that it is not performing that well (both resolution and chromatic aberration). After testing another three objectives of the same model, I swapped it for a 100x/1.4 and was much happier.  
   
Stan

Stanislav Vitha
Microscopy and Imaging Center
Texas A&M University

On Fri, 21 May 2021 10:03:57 +0000, Alison J. North <[hidden email]> wrote:

>Hi Abby,
>
>I don't know whether this will be useful information or not, but we recently purchased the 60X X-line objective for our DeltaVision, and I can confirm that the psfs look great and the chromatic aberrations are significantly better than with the original 60x/1.42 oil objective, even when using the Cargille oils (which result in noticeable chromatic dispersion in comparison to when using Olympus oil).  If the 100x X-line objective is anything like as good, then I really should be thinking about buying one myself - with the new 60x we no longer need to do any post-acquisition corrections.
>
>All the best,
>Alison
>
>Alison J. North, Ph.D.
>Senior Director of the Bio-Imaging Resource Center and Research Associate Professor
>The Rockefeller University
>1230 York Avenue
>New York
>NY 10065
>Tel. [phone] (office - direct line)
>https://www.rockefeller.edu/bioimaging/
>RRID:SCR_017791
>