movable objective nosepiece and resonant scanner on Zeiss multiphoton

[Sylvie Le Guyader]

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Hi Kate and others



I definitely agree that defining clearly the users' needs then getting a demo with their samples is a good place to start and a must to avoid huge disappointments.



I have questions regarding Zeiss multiphoton systems:

-          does anyone know if it is possible on a Zeiss multiphoton system to get a movable objective piece to accommodate different imaging angles during intravital imaging? If not, did anyone come up with their own solution?

-          Also I asked Zeiss if they have a resonant scanner and the answer was that they don't but that their new Fast is their solution. How to Fast compare to a resonant scanner? My understanding is that the detection with Fast is descanned so it must be less sensitive than a NDD solution with a resonant scanner. Does anyone have any experience with this?



thanks



Med vänlig hälsning / Best regards



Sylvie



@@@@@@@@@@@@@@@@@@@@@@@@

Sylvie Le Guyader, PhD

Live Cell Imaging Facility Manager

Karolinska Institutet- Bionut Dpt

Hälsovägen 7,

Novum, G lift, floor 6

14157 Huddinge

Sweden

mobile: [phone]

office: [phone]

LCI website





-----Original Message-----

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kate Luby-Phelps

Sent: den 16 mars 2017 16:37

To: [hidden email]

Subject: Re: commercially available multi-photon systems reviews



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Hi Rominy,



We have a Zeiss LSM780 with a Coherent Vision laser and a BiG two channel non-descanned detector on an upright AxioExaminer stand in our multiuser core and it has proved very useful. Based on about five years experience with multiphoton in a multiuser core facility here are some additional things to put into your equation:



1. First of all, what applications do you anticipate? If you need to go as deep as possible (approx. 1mm) then you will need objectives with long working distance, optimally water immersion if your samples are aqueous, and you might need to customize your lightpath for deepest possible imaging (not possible with every commercial vendor). I have known people to choose the vendor based on what objectives are offered and/or how accessible the lightpath is to modification.



2. If you are going to do live animal imaging in a variety of tissues, you might find that the biggest challenge is getting the animal on the stage in the correct orientation and immobilized. We opted to let the users solve those logistical problems themselves as we have many users with different requirements. With the availability of 3D printing or a good machine shop, custom solutions are within the reach of most investigators.



3. It can be difficult or impossible to excite far red probes like CY5 with NIR lasers. If that is important you might want to consider including an OPO or other solution for this issue.



4. I have always found that a extensive hands on demo with my own users' samples is helpful for uncovering unanticipated details that can help you with your decision.



Have fun shopping!



Kate

[Csúcs Gábor-3]

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Dear Slyvie,

Indeed the Zeiss FAST (which needs to be combined with Airy-scan) can reach the speeds of a resonant scanner. But you are correct - it is descanned, so you certainly loose signal. Furthermore, currently the Airy-scan/Fast is available only in 1-channel mode, multiple colors you need to acquire sequentially.

Greetings    Gabor

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Sylvie Le Guyader
Sent: Freitag, 17. März 2017 09:39
To: [hidden email]
Subject: movable objective nosepiece and resonant scanner on Zeiss multiphoton

*****
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 Kate and others



I definitely agree that defining clearly the users' needs then getting a demo with their samples is a good place to start and a must to avoid huge disappointments.



I have questions regarding Zeiss multiphoton systems:

-          does anyone know if it is possible on a Zeiss multiphoton system to get a movable objective piece to accommodate different imaging angles during intravital imaging? If not, did anyone come up with their own solution?

-          Also I asked Zeiss if they have a resonant scanner and the answer was that they don't but that their new Fast is their solution. How to Fast compare to a resonant scanner? My understanding is that the detection with Fast is descanned so it must be less sensitive than a NDD solution with a resonant scanner. Does anyone have any experience with this?



thanks



Med vänlig hälsning / Best regards



Sylvie



@@@@@@@@@@@@@@@@@@@@@@@@

Sylvie Le Guyader, PhD

Live Cell Imaging Facility Manager

Karolinska Institutet- Bionut Dpt

Hälsovägen 7,

Novum, G lift, floor 6

14157 Huddinge

Sweden

mobile: [phone]

office: [phone]

LCI website





-----Original Message-----

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kate Luby-Phelps

Sent: den 16 mars 2017 16:37

To: [hidden email]

Subject: Re: commercially available multi-photon systems reviews



*****

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 Rominy,



We have a Zeiss LSM780 with a Coherent Vision laser and a BiG two channel non-descanned detector on an upright AxioExaminer stand in our multiuser core and it has proved very useful. Based on about five years experience with multiphoton in a multiuser core facility here are some additional things to put into your equation:



1. First of all, what applications do you anticipate? If you need to go as deep as possible (approx. 1mm) then you will need objectives with long working distance, optimally water immersion if your samples are aqueous, and you might need to customize your lightpath for deepest possible imaging (not possible with every commercial vendor). I have known people to choose the vendor based on what objectives are offered and/or how accessible the lightpath is to modification.



2. If you are going to do live animal imaging in a variety of tissues, you might find that the biggest challenge is getting the animal on the stage in the correct orientation and immobilized. We opted to let the users solve those logistical problems themselves as we have many users with different requirements. With the availability of 3D printing or a good machine shop, custom solutions are within the reach of most investigators.



3. It can be difficult or impossible to excite far red probes like CY5 with NIR lasers. If that is important you might want to consider including an OPO or other solution for this issue.



4. I have always found that a extensive hands on demo with my own users' samples is helpful for uncovering unanticipated details that can help you with your decision.



Have fun shopping!



Kate