LSM 800 - 405 nm laser collimator

[ramachan]

*****
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.
*****

Dear All,

A colleague asked me to have a look at their LSM 800, where they face problems with huge axial shifts between 405 nm-channel and the rest of the channels (12 nm for 40x water dipping and even more with 20x). I had never seen an LSM 800 before and being familiar with LSM 780/880 or Leica SP8, I've expected it to be the matter or 405 nm collimator alignment. However, when seeing the actual setup, I've noticed there was a single fibre coming to the scan head and there was no control over 405 nm collimator in the software. Local Zeiss representatives claim it to be caused by the chromatic aberration if the lenses and propose an upgrade to higher-end lenses. While those higher-end lenses may be better in many ways, I am not convinced the proposed upgrade will entirely solve the problem. Can anybody familiar with the internal workings of an LSM 800 comment on this? Is there really a single collimator for all laser lines, in which case I wonder what can cause such huge shifts between the channels, or have I overlooked something?

Thanks! Best wishes,
Radek

Radek MACHAN, Ph.D. (Senior Research Fellow)
SCELSE Advanced Biofilm Imaging Facility<http://www.scelse.sg/Page/imaging-facility> Manager
Nanyang Technological University
#B2, 60 Nanyang Drive, SBS-01N-27
Singapore 637551


________________________________

CONFIDENTIALITY: This email is intended solely for the person(s) named and may be confidential and/or privileged. If you are not the intended recipient, please delete it, notify us and do not copy, use, or disclose its contents.
Towards a sustainable earth: Print only when necessary. Thank you.

[Thomas Pitta-2]

*****
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.
*****

This is most certainly an objective issue. You need a 405 corrected objective.

Sent from my iPhone

> On Sep 20, 2020, at 6:40 PM, Radek Machan (Dr) <[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.
> *****
>
> Dear All,
>
> A colleague asked me to have a look at their LSM 800, where they face problems with huge axial shifts between 405 nm-channel and the rest of the channels (12 nm for 40x water dipping and even more with 20x). I had never seen an LSM 800 before and being familiar with LSM 780/880 or Leica SP8, I've expected it to be the matter or 405 nm collimator alignment. However, when seeing the actual setup, I've noticed there was a single fibre coming to the scan head and there was no control over 405 nm collimator in the software. Local Zeiss representatives claim it to be caused by the chromatic aberration if the lenses and propose an upgrade to higher-end lenses. While those higher-end lenses may be better in many ways, I am not convinced the proposed upgrade will entirely solve the problem. Can anybody familiar with the internal workings of an LSM 800 comment on this? Is there really a single collimator for all laser lines, in which case I wonder what can cause such huge shifts between the channels, or have I overlooked something?
>
> Thanks! Best wishes,
> Radek
>
> Radek MACHAN, Ph.D. (Senior Research Fellow)
> SCELSE Advanced Biofilm Imaging Facility<http://www.scelse.sg/Page/imaging-facility> Manager
> Nanyang Technological University
> #B2, 60 Nanyang Drive, SBS-01N-27
> Singapore 637551
>
>
> ________________________________
>
> CONFIDENTIALITY: This email is intended solely for the person(s) named and may be confidential and/or privileged. If you are not the intended recipient, please delete it, notify us and do not copy, use, or disclose its contents.
> Towards a sustainable earth: Print only when necessary. Thank you.

[David Claypool]

*****
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 agree with Tom. You need an objective that is, also, axially well corrected for 405nm.

David Claypool

> On Sep 20, 2020, at 7:16 PM, Thomas Pitta <[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.
> *****
>
> This is most certainly an objective issue. You need a 405 corrected objective.
>
> Sent from my iPhone
>
>> On Sep 20, 2020, at 6:40 PM, Radek Machan (Dr) <[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.
>> *****
>>
>> Dear All,
>>
>> A colleague asked me to have a look at their LSM 800, where they face problems with huge axial shifts between 405 nm-channel and the rest of the channels (12 nm for 40x water dipping and even more with 20x). I had never seen an LSM 800 before and being familiar with LSM 780/880 or Leica SP8, I've expected it to be the matter or 405 nm collimator alignment. However, when seeing the actual setup, I've noticed there was a single fibre coming to the scan head and there was no control over 405 nm collimator in the software. Local Zeiss representatives claim it to be caused by the chromatic aberration if the lenses and propose an upgrade to higher-end lenses. While those higher-end lenses may be better in many ways, I am not convinced the proposed upgrade will entirely solve the problem. Can anybody familiar with the internal workings of an LSM 800 comment on this? Is there really a single collimator for all laser lines, in which case I wonder what can cause such huge shifts between the channels, or have I overlooked something?
>>
>> Thanks! Best wishes,
>> Radek
>>
>> Radek MACHAN, Ph.D. (Senior Research Fellow)
>> SCELSE Advanced Biofilm Imaging Facility<http://www.scelse.sg/Page/imaging-facility> Manager
>> Nanyang Technological University
>> #B2, 60 Nanyang Drive, SBS-01N-27
>> Singapore 637551
>>
>>
>> ________________________________
>>
>> CONFIDENTIALITY: This email is intended solely for the person(s) named and may be confidential and/or privileged. If you are not the intended recipient, please delete it, notify us and do not copy, use, or disclose its contents.
>> Towards a sustainable earth: Print only when necessary. Thank you.

[Tomek Węgierski]

*****
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.
*****

Dear Radek,
Just to clarify: do you really call a 12 nm axial shift between the 405 channel and others "huge"?
I hope this is some kind of typo, otherwise how should I refer to the axial shift I see on our LSM800?
BTW: in ZEN for LSM800 there is a way to introduce an axial offset between channels, so shifts can be corrected already during the acquisition. It does not solve the hardware problem but it may be helpful.
best,
Tomek

Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
http://www.iimcb.gov.pl/
________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Radek Machan (Dr) <[hidden email]>
Sent: Monday, September 21, 2020 12:39 AM
To: [hidden email] <[hidden email]>
Subject: LSM 800 - 405 nm laser collimator

*****
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.
*****

Dear All,

A colleague asked me to have a look at their LSM 800, where they face problems with huge axial shifts between 405 nm-channel and the rest of the channels (12 nm for 40x water dipping and even more with 20x). I had never seen an LSM 800 before and being familiar with LSM 780/880 or Leica SP8, I've expected it to be the matter or 405 nm collimator alignment. However, when seeing the actual setup, I've noticed there was a single fibre coming to the scan head and there was no control over 405 nm collimator in the software. Local Zeiss representatives claim it to be caused by the chromatic aberration if the lenses and propose an upgrade to higher-end lenses. While those higher-end lenses may be better in many ways, I am not convinced the proposed upgrade will entirely solve the problem. Can anybody familiar with the internal workings of an LSM 800 comment on this? Is there really a single collimator for all laser lines, in which case I wonder what can cause such huge shifts between the channels, or have I overlooked something?

Thanks! Best wishes,
Radek

Radek MACHAN, Ph.D. (Senior Research Fellow)
SCELSE Advanced Biofilm Imaging Facility<http://www.scelse.sg/Page/imaging-facility> Manager
Nanyang Technological University
#B2, 60 Nanyang Drive, SBS-01N-27
Singapore 637551


________________________________

CONFIDENTIALITY: This email is intended solely for the person(s) named and may be confidential and/or privileged. If you are not the intended recipient, please delete it, notify us and do not copy, use, or disclose its contents.
Towards a sustainable earth: Print only when necessary. Thank you.

[ramachan]

*****
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.
*****

Dear Tomek,

Yes, you are right there was a typo – should read microns. They did not attempt any super-resolution there.

Best,
Radek


Radek MACHAN, Ph.D. (Senior Research Fellow)

SCELSE Advanced Biofilm Imaging Facility<http://www.scelse.sg/Page/imaging-facility> Manager

Nanyang Technological University
#B2, 60 Nanyang Drive, SBS-01N-27
Singapore 637551

From: Tomek Węgierski<mailto:[hidden email]>
Sent: Monday, 21 September 2020 4:47 PM
To: [hidden email]<mailto:[hidden email]>
Subject: Re: LSM 800 - 405 nm laser collimator

*****
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.
*****

Dear Radek,
Just to clarify: do you really call a 12 nm axial shift between the 405 channel and others "huge"?
I hope this is some kind of typo, otherwise how should I refer to the axial shift I see on our LSM800?
BTW: in ZEN for LSM800 there is a way to introduce an axial offset between channels, so shifts can be corrected already during the acquisition. It does not solve the hardware problem but it may be helpful.
best,
Tomek

Tomasz Wegierski, PhD
International Institute of Molecular and Cell Biology
Trojdena 4, 02-109 Warsaw, POLAND
tel: [phone]
fax: [phone]
http://www.iimcb.gov.pl/
________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Radek Machan (Dr) <[hidden email]>
Sent: Monday, September 21, 2020 12:39 AM
To: [hidden email] <[hidden email]>
Subject: LSM 800 - 405 nm laser collimator

*****
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.
*****

Dear All,

A colleague asked me to have a look at their LSM 800, where they face problems with huge axial shifts between 405 nm-channel and the rest of the channels (12 nm for 40x water dipping and even more with 20x). I had never seen an LSM 800 before and being familiar with LSM 780/880 or Leica SP8, I've expected it to be the matter or 405 nm collimator alignment. However, when seeing the actual setup, I've noticed there was a single fibre coming to the scan head and there was no control over 405 nm collimator in the software. Local Zeiss representatives claim it to be caused by the chromatic aberration if the lenses and propose an upgrade to higher-end lenses. While those higher-end lenses may be better in many ways, I am not convinced the proposed upgrade will entirely solve the problem. Can anybody familiar with the internal workings of an LSM 800 comment on this? Is there really a single collimator for all laser lines, in which case I wonder what can cause such huge shifts between the channels, or have I overlooked something?

Thanks! Best wishes,
Radek

Radek MACHAN, Ph.D. (Senior Research Fellow)
SCELSE Advanced Biofilm Imaging Facility<http://www.scelse.sg/Page/imaging-facility> Manager
Nanyang Technological University
#B2, 60 Nanyang Drive, SBS-01N-27
Singapore 637551


________________________________

CONFIDENTIALITY: This email is intended solely for the person(s) named and may be confidential and/or privileged. If you are not the intended recipient, please delete it, notify us and do not copy, use, or disclose its contents.
Towards a sustainable earth: Print only when necessary. Thank you.