LSM880 filters for the Airyscan module

[Romin, Yevgeniy/Sloan Kettering Institute]

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Hello everybody

This is a question for those of you who has the LSM 880 with Airyscan module in your lab or core facility.  Has anyone dealt with swapping out the emission filters in the internal filterwheel that goes in front of the airyscan detector?  We are having considerable crosstalk issues when we try to do multi-channel airyscan imaging.  If anybody has come across this situation and can share how you dealt with it (what's the filter combination that you have used, etc), It will be much appreciated.

Thanks to all of you in advance,

Yevgeniy Romin

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[Krzic, Uros]

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Hi Yevgeniy,

 

Reports of crosstalk issues with Airyscan have been very rare so far. Crosstalk seems to become a problem only when i) dyes with closely overlapping spectra are imaged e.g. GFP-YFP or similar, ii) one or more excitation wavelengths do not match the dyes’ excitation peaks, which might lead to cross-excitation and iii) concentrations of different dyes are vastly different (an order of magnitude or more). In all other cases it is normally easy to separate the channels using the standard set of Airyscan emission filters, especially if a different filter settings are used for every channel (“Switch track every image”). Moreover, the LSM 880’s secondary beam splitter (SBS) can often be used to further optimize Airyscan’s detection spectrum.

 

Of course if this does not help, you can contact your local Zeiss service and ask them to replace the filters in the Airyscan filter wheel (perhaps with single-band filters) and/or filters in the SBS. They will advise you on the best combination of filters for your application and arrange the exchange.

 

With best regards from Munich,

Uros, Carl Zeiss Microscopy

 

 

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of [hidden email]
Sent: Dienstag, 6. September 2016 16:17
To: [hidden email]
Subject: LSM880 filters for the Airyscan module

 

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Hello everybody

 

This is a question for those of you who has the LSM 880 with Airyscan module in your lab or core facility.  Has anyone dealt with swapping out the emission filters in the internal filterwheel that goes in front of the airyscan detector?  We are having considerable crosstalk issues when we try to do multi-channel airyscan imaging.  If anybody has come across this situation and can share how you dealt with it (what's the filter combination that you have used, etc), It will be much appreciated.

 

Thanks to all of you in advance,

 

Yevgeniy Romin

=====================================================================

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[Anne Vaahtokari]

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Hi Yevgeniy,

 

Have you tried linear unmixing to get rid of the crosstalk? That works well for us.

 

Anne

 

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of [hidden email]
Sent: Tuesday, September 06, 2016 8:17 AM
To: [hidden email]
Subject: LSM880 filters for the Airyscan module

 

***** 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. *****

Hello everybody

 

This is a question for those of you who has the LSM 880 with Airyscan module in your lab or core facility.  Has anyone dealt with swapping out the emission filters in the internal filterwheel that goes in front of the airyscan detector?  We are having considerable crosstalk issues when we try to do multi-channel airyscan imaging.  If anybody has come across this situation and can share how you dealt with it (what's the filter combination that you have used, etc), It will be much appreciated.

 

Thanks to all of you in advance,

 

Yevgeniy Romin

=====================================================================

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[Deanne Veronica Catmull]

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I don't have an LSM 880 but we were just given a seminar on the functionality and capabilities of the system here yesterday as one has been installed on campus. My belief from the presentation given yesterday, that you can perform instant unmixing of flourophores with simultaneous acquisition using the airyscan in the 880 (but only with sequential scanning in the 800). You shouldn't get cross talk issues. There are plenty of detectors and beam path options that you could set up to avoid this.

Kind regards,

Deanne.

Deanne Catmull | Research Assistant and HSR
Melbourne Dental School | Faculty of Medicine, Dentistry and Health Sciences

Level 6, 720 Swanston St
The University of Melbourne, Victoria 3010 Australia
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From: Confocal Microscopy List <[hidden email]> on behalf of Anne Vaahtokari <[hidden email]>
Sent: Tuesday, 13 September 2016 1:34:47 AM
To: [hidden email]
Subject: Re: LSM880 filters for the Airyscan module

 

***** 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 Yevgeniy,

 

Have you tried linear unmixing to get rid of the crosstalk? That works well for us.

 

Anne

 

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of [hidden email]
Sent: Tuesday, September 06, 2016 8:17 AM
To: [hidden email]
Subject: LSM880 filters for the Airyscan module

 

***** 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. *****

Hello everybody

 

This is a question for those of you who has the LSM 880 with Airyscan module in your lab or core facility.  Has anyone dealt with swapping out the emission filters in the internal filterwheel that goes in front of the airyscan detector?  We are having considerable crosstalk issues when we try to do multi-channel airyscan imaging.  If anybody has come across this situation and can share how you dealt with it (what's the filter combination that you have used, etc), It will be much appreciated.

 

Thanks to all of you in advance,

 

Yevgeniy Romin

=====================================================================

Please note that this e-mail and any files transmitted from
Memorial Sloan Kettering Cancer Center may be privileged, confidential,
and protected from disclosure under applicable law. If the reader of
this message is not the intended recipient, or an employee or agent
responsible for delivering this message to the intended recipient,
you are hereby notified that any reading, dissemination, distribution,
copying, or other use of this communication or any of its attachments
is strictly prohibited. If you have received this communication in
error, please notify the sender immediately by replying to this message
and deleting this message, any attachments, and all copies and backups
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