Deep tissue imaging with spinning disc - comparison with other confocal techniques

[Jan Pala]

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Dear listservers,

looking for some application techniques with deep tissue imaging I found
this article:
http://neuronet.jp/pdf/O_101.pdf
Journal of Integrative Neuroscience, Vol. 10, No. 1 (2011) 121–129
DOI: 10.1142/S0219635211002658
NIPKOW CONFOCAL IMAGING FROM DEEP BRAIN TISSUES
YUJI TAKAHARA, NORIO MATSUKI and YUJI IKEGAYA

where the authors describe advantages of spinning disc in comparison with
standard confocal laser scanning microscope and two-photon microscope.

What do  you think about presented results and experimental conditions? For
me, it is quite strange, see part 3.1:
1. the laser power was fixed at 5 mW in all microscope systems - for CLSM
the power is too high, for multiphoton too low. There is no description
about the way the power was measured and set
2. Real Z-resolution for all the systems
3. Results: CLSM penetration depth 80 microns, spinning disc penetration
depth 150 microns and multiphoton penetration depth 200 microns - it is
quite unbelieveable to see almost double penetration depth in between
spinning disc and CLSM.

Thank you in advance.

Best regards,
Jan Pala

[Arne Seitz]

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Dear Jan,

you are asking our opinion on an article. But the way you are asking is already guiding into a specific direction. I personally do not like what you are trying to do (for me it goes into the direction of paper "bashing"). If you have questions/doubts regarding this technique or the results you should address it to the authors of the paper. Maybe it is just a misunderstanding (I cannot download the pdf properly so it is hard to judge) which could be ruled out by looking at the original data.

Just my 2c.

Best regards
Arne


---------------------------------------------------------------
Arne Seitz
Head of Bioimaging and Optics Platform (PT-BIOP)
Ecole Polytechnique Fédérale de Lausanne (EPFL)
Faculty of Life Sciences
Station 15, AI 0241
CH-1015 Lausanne

Phone: [phone]
Fax:      [phone]
http://biop.epfl.ch/
---------------------------------------------------------------

> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]] On Behalf Of Jan Pala
> Sent: mercredi 16 mai 2012 12:50
> To: [hidden email]
> Subject: Deep tissue imaging with spinning disc - comparison with other
> confocal techniques
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear listservers,
>
> looking for some application techniques with deep tissue imaging I found this
> article:
> http://neuronet.jp/pdf/O_101.pdf
> Journal of Integrative Neuroscience, Vol. 10, No. 1 (2011) 121-129
> DOI: 10.1142/S0219635211002658
> NIPKOW CONFOCAL IMAGING FROM DEEP BRAIN TISSUES YUJI TAKAHARA,
> NORIO MATSUKI and YUJI IKEGAYA
>
> where the authors describe advantages of spinning disc in comparison with
> standard confocal laser scanning microscope and two-photon microscope.
>
> What do  you think about presented results and experimental conditions?
> For me, it is quite strange, see part 3.1:
> 1. the laser power was fixed at 5 mW in all microscope systems - for CLSM the
> power is too high, for multiphoton too low. There is no description about the
> way the power was measured and set 2. Real Z-resolution for all the systems
> 3. Results: CLSM penetration depth 80 microns, spinning disc penetration
> depth 150 microns and multiphoton penetration depth 200 microns - it is
> quite unbelieveable to see almost double penetration depth in between
> spinning disc and CLSM.
>
> Thank you in advance.
>
> Best regards,
> Jan Pala

[Jan Pala]

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Dear Arne,

you are right that from the way my points were written it is somehow guided
- pardon for that. I was quit surprised from presented results, i.e.
spinning disc is almost twice better than standard CLSM and used laser
power, that I was not enough careful with proper way of asking for opinions
and ideas from members of confocal list server. Sorry for any inconvenience...

If there is any problem with downloading the article I can send it via email.

Best regards,
Jan Pala

[George McNamara]

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Hi Jan,

The reason to use confocal - whether spinning disk or laser scanning -
is to block the out of focus light. In their Figure 1, two-photon did an
excellent job through the entire 200 um range shown (they should have
kept going!).

Confocal laser scanning cut out at ~80 um as the authors mention.

The Yokogawa shows a couple of in focus, or maybe "not quite in focus"
features at 100 um, but everything at and below 120 um looks blurry =
out of focus.

Bottom line: Multiphoton excitation wins again.

George
p.s. the authors failed to show whether the MP excitation wavelength was
optimal for EGFP or any of the dyes evaluated. They also did not state
the pinhole size for MPEF or whether they used NDD path for MPEF. For
spinning disk, they used an iXonEM+ DV897 EMCCD, but did not state the
EM gain or even if they used the EM gain register (probably).


On 5/16/2012 6:49 AM, Jan Pala wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear listservers,
>
> looking for some application techniques with deep tissue imaging I found
> this article:
> http://neuronet.jp/pdf/O_101.pdf
> Journal of Integrative Neuroscience, Vol. 10, No. 1 (2011) 121–129
> DOI: 10.1142/S0219635211002658
> NIPKOW CONFOCAL IMAGING FROM DEEP BRAIN TISSUES
> YUJI TAKAHARA, NORIO MATSUKI and YUJI IKEGAYA
>
> where the authors describe advantages of spinning disc in comparison with
> standard confocal laser scanning microscope and two-photon microscope.
>
> What do  you think about presented results and experimental conditions? For
> me, it is quite strange, see part 3.1:
> 1. the laser power was fixed at 5 mW in all microscope systems - for CLSM
> the power is too high, for multiphoton too low. There is no description
> about the way the power was measured and set
> 2. Real Z-resolution for all the systems
> 3. Results: CLSM penetration depth 80 microns, spinning disc penetration
> depth 150 microns and multiphoton penetration depth 200 microns - it is
> quite unbelieveable to see almost double penetration depth in between
> spinning disc and CLSM.
>
> Thank you in advance.
>
> Best regards,
> Jan Pala
>
>    

[Sebastian Rhode]

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Hi Jan,

since our company manufactures both kind of systems, spinning disc confocal
and 2-Photon systems, just my opinion on this paper.

What I have seen so far during our tests, shows a clear advantage of 2PM if
depth penetration is your primary(!) goal. If it comes to speed, spinning
disc confocal (SDC) systems have a clear advantage. And in terms of cell
viability, 2PM and SDC are better than a conventional LSM microscope.

What puzzles me a bit, is the comparison between the penetration depth of
the SDC and LSM. There are several points (some of them already mentioned by
George here), which makes it difficult for me to really compare the results.

But nevertheless the findings are quite interesting, even if there are open
questions to answer. But I would be really interested in the explanation
about the observed results.

Dr. Sebastian Rhode
Project Manager Research & Development      

TILL Photonics GmbH
an FEI Company

[Jan Pala]

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Dear Sebastian,

I agree with George and you. I still do not understand the excitation
comparison - based on your experience, is there somehow comparable the
excitation laser power in between different confocal principles (5 mW) as
presented in the paper?

For sure, I am also very interested in some deep explanation and analysis
of observed and presented results.

Thank you.

Best regards,
Jan

On Fri, May 18, 2012 at 9:10 AM, Sebastian Rhode <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Jan,
>
> since our company manufactures both kind of systems, spinning disc confocal
> and 2-Photon systems, just my opinion on this paper.
>
> What I have seen so far during our tests, shows a clear advantage of 2PM if
> depth penetration is your primary(!) goal. If it comes to speed, spinning
> disc confocal (SDC) systems have a clear advantage. And in terms of cell
> viability, 2PM and SDC are better than a conventional LSM microscope.
>
> What puzzles me a bit, is the comparison between the penetration depth of
> the SDC and LSM. There are several points (some of them already mentioned
> by
> George here), which makes it difficult for me to really compare the
> results.
>
> But nevertheless the findings are quite interesting, even if there are open
> questions to answer. But I would be really interested in the explanation
> about the observed results.
>
> Dr. Sebastian Rhode
> Project Manager Research & Development
>
> TILL Photonics GmbH
> an FEI Company
>

--
http://palic.ho-vsetin.com
http://www.mountainski.cz
[hidden email]
icq: 133-783-208

[George McNamara]

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MPEF can also be fast with resonant scanner (ex. Leica) or TriMScope
(LaVision Biotec, which uses array of spots and camera ... more spots,
less depth).

For example of fast resonant scanner with confocal see:

High-resolution, noninvasive longitudinal live imaging of immune
responses. </pubmed/21768391> Abdulreda MH, Faleo G, Molano RD,
Lopez-Cabezas M, Molina J, Tan Y, Echeverria OA, Zahr-Akrawi E,
Rodriguez-Diaz R, Edlund PK, Leibiger I, Bayer AL, Perez V, Ricordi C,
Caicedo A, Pileggi A, Berggren PO. Proc Natl Acad Sci U S A. 2011 Aug
2;108(31):12863-8. Epub 2011 Jul 18. PMID: 21768391.

This paper just used confocal lasers. If someone will send us money for
four HyD detectors NDD module, I'll get my colleagues to use our
Chameleon Ultra II more often.




On 5/18/2012 3:10 AM, Sebastian Rhode wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Jan,
>
> since our company manufactures both kind of systems, spinning disc confocal
> and 2-Photon systems, just my opinion on this paper.
>
> What I have seen so far during our tests, shows a clear advantage of 2PM if
> depth penetration is your primary(!) goal. If it comes to speed, spinning
> disc confocal (SDC) systems have a clear advantage. And in terms of cell
> viability, 2PM and SDC are better than a conventional LSM microscope.
>
> What puzzles me a bit, is the comparison between the penetration depth of
> the SDC and LSM. There are several points (some of them already mentioned by
> George here), which makes it difficult for me to really compare the results.
>
> But nevertheless the findings are quite interesting, even if there are open
> questions to answer. But I would be really interested in the explanation
> about the observed results.
>
> Dr. Sebastian Rhode
> Project Manager Research&  Development
>
> TILL Photonics GmbH
> an FEI Company
>
>    

[Mark Cannell]

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The improved penetration of SDC compared to confocal is almost certainly the wider pinhole of the SDC compared to confocal IMHO. As a shameless self citation, I think that the penetration depth issue and effect of pinhoie etc. are quite well illustrated in our paper:

Soeller C, Cannell MB. Two-photon microscopy: Imaging in scattering samples and three-dimensionally resolved flash photolysis. Microsc. Res. Tech. 1999;47(3):182–95.


Note the effect of opening the pinhole fully in our Fig. 4 and the relative performance loss due to closing the pinhole with depth (highly non-linear with essentially no signal beyond 50 um in this sample). In this strongly scattering sample (cheese), 1% signal was obtained at 45 um with a pinhole,  opening the pinhole increased the signal ~10x and using non-descanned detection increased the signal ~3x more. The method used 2P excitation in all cases so that we could examine the detector side performance -so the differences would be even greater for 1P where scattering also decreases excitation intensity at the focal spot ...

Cheers Mark


On 18/05/2012, at 8:10 AM, Sebastian Rhode wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Jan,
>
> since our company manufactures both kind of systems, spinning disc confocal
> and 2-Photon systems, just my opinion on this paper.
>
> What I have seen so far during our tests, shows a clear advantage of 2PM if
> depth penetration is your primary(!) goal. If it comes to speed, spinning
> disc confocal (SDC) systems have a clear advantage. And in terms of cell
> viability, 2PM and SDC are better than a conventional LSM microscope.
>
> What puzzles me a bit, is the comparison between the penetration depth of
> the SDC and LSM. There are several points (some of them already mentioned by
> George here), which makes it difficult for me to really compare the results.
>
> But nevertheless the findings are quite interesting, even if there are open
> questions to answer. But I would be really interested in the explanation
> about the observed results.
>
> Dr. Sebastian Rhode
> Project Manager Research & Development      
>
> TILL Photonics GmbH
> an FEI Company

[Sebastian Rhode]

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Hi Mark,

shame on me, that did not really read the paper. So when using a 20X NA0.95
objective, the required pinhole should have been ~13 micron in diameter
instead of the used 50 microns. We use 30 micron right now a I have not
looked really deep into our brain slices using our Andromeda SDC system.

Cheers,

Sebi

[David Baddeley]

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Mark beat me to it in suggesting differences in pinhole size, which is probably the bulk of the explanation. I think that there is also another factor contributing, namely that defoccused spinning disk illumination looks rather like widefield illumination, so you might expect more signal (but not more contrast) at depth with a spinning disk system even if the pinholes were equivalent.

Cheers,
David


------------------------------
On Sat, May 19, 2012 1:32 AM NZST Mark Cannell wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>The improved penetration of SDC compared to confocal is almost certainly the wider pinhole of the SDC compared to confocal IMHO. As a shameless self citation, I think that the penetration depth issue and effect of pinhoie etc. are quite well illustrated in our paper:
>
>Soeller C, Cannell MB. Two-photon microscopy: Imaging in scattering samples and three-dimensionally resolved flash photolysis. Microsc. Res. Tech. 1999;47(3):182–95.
>
>
>Note the effect of opening the pinhole fully in our Fig. 4 and the relative performance loss due to closing the pinhole with depth (highly non-linear with essentially no signal beyond 50 um in this sample). In this strongly scattering sample (cheese), 1% signal was obtained at 45 um with a pinhole,  opening the pinhole increased the signal ~10x and using non-descanned detection increased the signal ~3x more. The method used 2P excitation in all cases so that we could examine the detector side performance -so the differences would be even greater for 1P where scattering also decreases excitation intensity at the focal spot ...
>
>Cheers Mark
>
>
>On 18/05/2012, at 8:10 AM, Sebastian Rhode wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hi Jan,
>>
>> since our company manufactures both kind of systems, spinning disc confocal
>> and 2-Photon systems, just my opinion on this paper.
>>
>> What I have seen so far during our tests, shows a clear advantage of 2PM if
>> depth penetration is your primary(!) goal. If it comes to speed, spinning
>> disc confocal (SDC) systems have a clear advantage. And in terms of cell
>> viability, 2PM and SDC are better than a conventional LSM microscope.
>>
>> What puzzles me a bit, is the comparison between the penetration depth of
>> the SDC and LSM. There are several points (some of them already mentioned by
>> George here), which makes it difficult for me to really compare the results.
>>
>> But nevertheless the findings are quite interesting, even if there are open
>> questions to answer. But I would be really interested in the explanation
>> about the observed results.
>>
>> Dr. Sebastian Rhode
>> Project Manager Research & Development      
>>
>> TILL Photonics GmbH
>> an FEI Company

[Julio Vazquez]

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Hi Jan,

Comparisons between systems have to be done very carefully to be really meaningful, since there are so many variables between instruments. Even between similar types of instruments, there can be substantial differences, the more so as the instruments get more complex. A slightly misaligned pinhole can send your confocal performance and image quality down the drain, and even 1 Airy unit may mean different things on different microscopes.  Just yesterday I was reading a brochure about one of the major vendor's new confocal, where they describe their hexagonal pinhole, which at 1 Airy units gives (according to the brochure) 30% more transmission than a square pinhole. The term pinhole tends to evoque a little circle in my mind, so I often forget that pinholes in confocals can be square, circular, or hexagonal. In the example above, clearly, the manufacturer was comparing the light throughput of an hexagonal pinhole with a square pinhole where the Airy unit would correspond to the diagonal. However, they don't say that their hexagonal pinhole would transmit less light than a circular pinhole, or in the case of square pinholes, 1 Airy Unit may be defined as the length of the square, in which case, the square pinhole still wins (with the hexagonal pinhole the Airy unit is defined as the largest diameter, i.e. the 1 Airy hexagon fits inside a 1 Airy circle). Anyway, this is just to illustrate that a trivial difference such as the geometry of the pinhole can result in a 20-30% difference in efficiency between two systems, all other things being equal (which they rarely are). Another trivial example, with one of our confocals, using to alternate (and theoretically functionally similar) dichroics can give as much as 20-30% difference in image intensity.

Back to the paper anyway, If you look at Figure 1, you'll see that the starting image (40 microns deep)  is about 2.5 times brighter with Nipkow than with conventional confocal (you can measure it with imagej, for what it is worth on a reproduction). If you crank up the contrast, you will see signal in the conventional confocal images down to 120-140 microns. How would those compare to the Nipkow system if the confocal image had been optimized, and initial brightness of the various systems had been equalized? who knows....

As you point out, I am also concerned by the 5 mW laser power used for all instruments. Besides the fact that we do not know where this power was measured, it is clear that this is way too much power to use on a conventional confocal (we typically use around 0.1 mW, definitely less than 0.5 mW, and I don't know too many samples that could take much imaging at 5 mW). On the other hand, it is probably a reasonable amount of power for a spinning disk (or widefield), and maybe on the low end for two-photon, but can't really say. In any event, if I were to collect a 200 micron stack on our point scanning confocal at 5 mW laser power, I would proably see massive bleaching after the first few sections. This could be what the authors are observing. Again, hard to know.

You may also notice that the fields of view shown in Fig 1 for all three systems are identical. Would the successive imaging of the same region on different microscopes not bleach the sample, which would obviously put the first instrument at an advantage? In this case, which images were acquired first, and which were acquired last?

--
Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA 98109-1024

http://www.fhcrc.org


----- Original Message -----
From: "Jan Pala" <[hidden email]>
To: [hidden email]
Sent: Friday, May 18, 2012 3:19:08 AM
Subject: Re: Deep tissue imaging with spinning disc - comparison with other confocal techniques

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Dear Sebastian,

I agree with George and you. I still do not understand the excitation
comparison - based on your experience, is there somehow comparable the
excitation laser power in between different confocal principles (5 mW) as
presented in the paper?

For sure, I am also very interested in some deep explanation and analysis
of observed and presented results.

Thank you.

Best regards,
Jan

On Fri, May 18, 2012 at 9:10 AM, Sebastian Rhode <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi Jan,
>
> since our company manufactures both kind of systems, spinning disc confocal
> and 2-Photon systems, just my opinion on this paper.
>
> What I have seen so far during our tests, shows a clear advantage of 2PM if
> depth penetration is your primary(!) goal. If it comes to speed, spinning
> disc confocal (SDC) systems have a clear advantage. And in terms of cell
> viability, 2PM and SDC are better than a conventional LSM microscope.
>
> What puzzles me a bit, is the comparison between the penetration depth of
> the SDC and LSM. There are several points (some of them already mentioned
> by
> George here), which makes it difficult for me to really compare the
> results.
>
> But nevertheless the findings are quite interesting, even if there are open
> questions to answer. But I would be really interested in the explanation
> about the observed results.
>
> Dr. Sebastian Rhode
> Project Manager Research & Development
>
> TILL Photonics GmbH
> an FEI Company
>

--
http://palic.ho-vsetin.com
http://www.mountainski.cz
[hidden email]
icq: 133-783-208

[Arvydas Matiukas]

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Dear list:
 
We just started working with Nikon C2 confocal. It is controlled by
NIS elements v4.0 software.  I wonder what is the best way
to allow multiple users  to keep independent imaging configurations.
It seems that just creating new Windows accounts but using the same
Nikon user name loads the same configuration for all users.
 
I had a perfect  solution for our Zeiss LSM 510 system where users
had separate windows accounts and separate configuration files
that were automatically loaded for corresponding user. It is not clear
what the NIS elements login is for and manual tells nothing about it.
 
Please share your experience how you addressed this multiple user problem
for Nikon sofware. Our local representative does not have  a solution yet.
 
 
Thank you,
Arvydas

 
 
 
Arvydas Matiukas, Ph.D.
Director of Confocal&Two-Photon Core
Department of Pharmacology
SUNY Upstate Medical University
766 Irving Ave., WH 3167
Syracuse, NY 13210
tel.: [phone]
fax: [phone]
email: [hidden email]

[Sylvie Le Guyader]

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Hi Arvydas

We use NIS accounts.

Create a single Windows account (admin rights) shared by all users. Then in the options you can create 1 NIS user accounts with common rights per user (don't forget to create 1 admin account for yourself and 1 for Nikon). You can set the common rights to what you want (not being able to modify the shared configs for example). Each user will get their private configurations. BEWARE: the first time you create a configuration in a new user account, it will be set to reloading the objective as well. All you need to do is untick the objective reload in the first config and the following one will have it unticked.

You then have access to all sorts of usage statistics per user.

Hope that helps

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader
Live Cell Imaging Unit
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14183 Huddinge
Sweden
office: [phone]
LCI room: [phone]
mobile: [phone]

> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]] On Behalf Of Arvydas
> Matiukas
> Sent: 24 May 2012 23:41
> To: [hidden email]
> Subject: Multiple users on Nikon Elements
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear list:
>
> We just started working with Nikon C2 confocal. It is controlled by NIS elements
> v4.0 software.  I wonder what is the best way to allow multiple users  to keep
> independent imaging configurations.
> It seems that just creating new Windows accounts but using the same Nikon user
> name loads the same configuration for all users.
>
> I had a perfect  solution for our Zeiss LSM 510 system where users had separate
> windows accounts and separate configuration files that were automatically loaded
> for corresponding user. It is not clear what the NIS elements login is for and
> manual tells nothing about it.
>
> Please share your experience how you addressed this multiple user problem for
> Nikon sofware. Our local representative does not have  a solution yet.
>
>
> Thank you,
> Arvydas
>
>
>
>
> Arvydas Matiukas, Ph.D.
> Director of Confocal&Two-Photon Core
> Department of Pharmacology
> SUNY Upstate Medical University
> 766 Irving Ave., WH 3167
> Syracuse, NY 13210
> tel.: [phone]
> fax: [phone]
> email: [hidden email]

[Arvydas Matiukas]

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Hi Sylvie,

I wonder if you have been using a "time logging function for imaging
facility managers" available
on NIS elements v 4.0 and if
you have a document/manual for it.

Thanks,
Arvydas

Arvydas Matiukas, Ph.D.
Director of Confocal&Two-Photon Imaging Core Facility
Department of Pharmacology
SUNY Upstate Medical University
766 Irving Ave., WH 3159
Syracuse, NY 13210
tel.: [phone]
fax: [phone]
email: [hidden email]

>>> Sylvie Le Guyader <[hidden email]> 05/25/12 04:15 AM >>>
*****
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Hi Arvydas

We use NIS accounts.

Create a single Windows account (admin rights) shared by all users. Then
in the options you can create 1 NIS user accounts with common rights per
user (don't forget to create 1 admin account for yourself and 1 for
Nikon). You can set the common rights to what you want (not being able
to modify the shared configs for example). Each user will get their
private configurations. BEWARE: the first time you create a
configuration in a new user account, it will be set to reloading the
objective as well. All you need to do is untick the objective reload in
the first config and the following one will have it unticked.

You then have access to all sorts of usage statistics per user.

Hope that helps

Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader
Live Cell Imaging Unit
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14183 Huddinge
Sweden
office: [phone]
LCI room: [phone]
mobile: [phone]

> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[hidden email]] On Behalf Of Arvydas
> Matiukas
> Sent: 24 May 2012 23:41
> To: [hidden email]
> Subject: Multiple users on Nikon Elements
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear list:
>
> We just started working with Nikon C2 confocal. It is controlled by

NIS elements
> v4.0 software.  I wonder what is the best way to allow multiple users
to keep
> independent imaging configurations.
> It seems that just creating new Windows accounts but using the same
Nikon user
> name loads the same configuration for all users.
>
> I had a perfect  solution for our Zeiss LSM 510 system where users had
separate
> windows accounts and separate configuration files that were
automatically loaded
> for corresponding user. It is not clear what the NIS elements login is
for and
> manual tells nothing about it.
>
> Please share your experience how you addressed this multiple user
problem for

> Nikon sofware. Our local representative does not have  a solution yet.
>
>
> Thank you,
> Arvydas
>
>
>
>
> Arvydas Matiukas, Ph.D.
> Director of Confocal&Two-Photon Core
> Department of Pharmacology
> SUNY Upstate Medical University
> 766 Irving Ave., WH 3167
> Syracuse, NY 13210
> tel.: [phone]
> fax: [phone]
> email: [hidden email]

[Kees van der Oord]

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To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Dear Arvydas,

Usage is logged in 'C:\ProgramData\Laboratory Imaging\Platform\Statistics.db'.
When NIS is not running, you can import this database in e.g. Access.
Nikon Instruments Inc service will update you with the instructions manuals
about this functionality.

WR,

Kees