Laser-based AF for Nikon Ti-S

[darren.thomson]

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Hello Listserv,

We've been told by Nikon that no hardware based AutoFocus system is compatible with our Nikon Ti-S body and that we have to acquire a new microscope. Has anyone had luck, or suggest alternatives? Software based AF won't do for our throughput applications.

Many thanks for your help,

Best,

Darren Thomson
Manchester Fungal Infection Group
University of Manchester

[Kyle Michael Douglass]

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Hi Darren,

I don't know the specifics of the Ti-S, but if it uses an oil objective with NA higher than your mounting medium, then you could try the pgFocus open hardware system from Karl Bellvé.

http://big.umassmed.edu/wiki/index.php/PgFocus

You will need a stage or objective scanner that accepts a 0 - 10 V analog control signal and to setup the IR reference beam optics yourself.

I use this for focus lock on a 60x oil widefield setup and can maintain focus to within 5 - 10 nm standard deviation. It can sometimes be tricky to scan the stage with the focus locked, but otherwise it works well.

Cheers,
Kyle


Dr. Kyle M. Douglass
Post-doctoral Researcher
EPFL - The Laboratory of Experimental Biophysics
http://leb.epfl.ch/
http://kmdouglass.github.io

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Darren Thomson [[hidden email]]
Sent: Tuesday, September 5, 2017 11:43 AM
To: [hidden email]
Subject: Laser-based AF for Nikon Ti-S

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*****

Hello Listserv,

We've been told by Nikon that no hardware based AutoFocus system is compatible with our Nikon Ti-S body and that we have to acquire a new microscope. Has anyone had luck, or suggest alternatives? Software based AF won't do for our throughput applications.

Many thanks for your help,

Best,

Darren Thomson
Manchester Fungal Infection Group
University of Manchester

[Kyle Michael Douglass]

*****
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*****

My apologies: I meant to say that the objective NA should be higher than the refractive index of the mounting medium so that the laser beam may be totally internally reflected from the coverslip/medium interface.

Kyle

Dr. Kyle M. Douglass
Post-doctoral Researcher
EPFL - The Laboratory of Experimental Biophysics
http://leb.epfl.ch/
http://kmdouglass.github.io

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Kyle Michael Douglass [[hidden email]]
Sent: Tuesday, September 5, 2017 12:14 PM
To: [hidden email]
Subject: Re: Laser-based AF for Nikon Ti-S

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*****

Hi Darren,

I don't know the specifics of the Ti-S, but if it uses an oil objective with NA higher than your mounting medium, then you could try the pgFocus open hardware system from Karl Bellvé.

http://big.umassmed.edu/wiki/index.php/PgFocus

You will need a stage or objective scanner that accepts a 0 - 10 V analog control signal and to setup the IR reference beam optics yourself.

I use this for focus lock on a 60x oil widefield setup and can maintain focus to within 5 - 10 nm standard deviation. It can sometimes be tricky to scan the stage with the focus locked, but otherwise it works well.

Cheers,
Kyle


Dr. Kyle M. Douglass
Post-doctoral Researcher
EPFL - The Laboratory of Experimental Biophysics
http://leb.epfl.ch/
http://kmdouglass.github.io

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Darren Thomson [[hidden email]]
Sent: Tuesday, September 5, 2017 11:43 AM
To: [hidden email]
Subject: Laser-based AF for Nikon Ti-S

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

Hello Listserv,

We've been told by Nikon that no hardware based AutoFocus system is compatible with our Nikon Ti-S body and that we have to acquire a new microscope. Has anyone had luck, or suggest alternatives? Software based AF won't do for our throughput applications.

Many thanks for your help,

Best,

Darren Thomson
Manchester Fungal Infection Group
University of Manchester

[Seamus Holden-2]

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The ASI CRISP autofocus system would probably work. It can be used to drive one of their Z piezo top plates or any generic Z-drive that accepts an input control voltage.

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Darren Thomson
Sent: 05 September 2017 10:43
To: [hidden email]
Subject: Laser-based AF for Nikon Ti-S

*****
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*****

Hello Listserv,

We've been told by Nikon that no hardware based AutoFocus system is compatible with our Nikon Ti-S body and that we have to acquire a new microscope. Has anyone had luck, or suggest alternatives? Software based AF won't do for our throughput applications.

Many thanks for your help,

Best,

Darren Thomson
Manchester Fungal Infection Group
University of Manchester

[darren.thomson]

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Thanks Kyle,

We will routinely use dry 20x/40x 0.7-0.9NA objectives.

Darren

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kyle Michael Douglass
Sent: 05 September 2017 11:20
To: [hidden email]
Subject: Re: Laser-based AF for Nikon Ti-S

*****
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*****

My apologies: I meant to say that the objective NA should be higher than the refractive index of the mounting medium so that the laser beam may be totally internally reflected from the coverslip/medium interface.

Kyle

Dr. Kyle M. Douglass
Post-doctoral Researcher
EPFL - The Laboratory of Experimental Biophysics http://leb.epfl.ch/ http://kmdouglass.github.io

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Kyle Michael Douglass [[hidden email]]
Sent: Tuesday, September 5, 2017 12:14 PM
To: [hidden email]
Subject: Re: Laser-based AF for Nikon Ti-S

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

Hi Darren,

I don't know the specifics of the Ti-S, but if it uses an oil objective with NA higher than your mounting medium, then you could try the pgFocus open hardware system from Karl Bellvé.

http://big.umassmed.edu/wiki/index.php/PgFocus

You will need a stage or objective scanner that accepts a 0 - 10 V analog control signal and to setup the IR reference beam optics yourself.

I use this for focus lock on a 60x oil widefield setup and can maintain focus to within 5 - 10 nm standard deviation. It can sometimes be tricky to scan the stage with the focus locked, but otherwise it works well.

Cheers,
Kyle


Dr. Kyle M. Douglass
Post-doctoral Researcher
EPFL - The Laboratory of Experimental Biophysics http://leb.epfl.ch/ http://kmdouglass.github.io

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Darren Thomson [[hidden email]]
Sent: Tuesday, September 5, 2017 11:43 AM
To: [hidden email]
Subject: Laser-based AF for Nikon Ti-S

*****
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 Listserv,

We've been told by Nikon that no hardware based AutoFocus system is compatible with our Nikon Ti-S body and that we have to acquire a new microscope. Has anyone had luck, or suggest alternatives? Software based AF won't do for our throughput applications.

Many thanks for your help,

Best,

Darren Thomson
Manchester Fungal Infection Group
University of Manchester

[Kyle Michael Douglass]

*****
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Post images on http://www.imgur.com and include the link in your posting.
*****

Hi Darren,

    We will routinely use dry 20x/40x 0.7-0.9NA objectives.

In that case I can also second Seamus's suggestion for the ASI CRISP. It uses an IR LED and Foucalt's method. Since you're using a dry objective, you should get a nice and bright reflection from the slide. It's more difficult with immersion objectives.

I've used the CRISP in conjunction with a PI piezo scanner. The only problems I encountered were related to software bugs in the calibration routine and a limited LED lifetime in a 5% CO2 humidified environment. ASI sent us a replacement LED and, if I recall correctly, fixed the software bugs with a firmware update, so don't be afraid to talk to them.

Cheers,
Kyle

Dr. Kyle M. Douglass
Post-doctoral Researcher
EPFL - The Laboratory of Experimental Biophysics
http://leb.epfl.ch/
http://kmdouglass.github.io

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Darren Thomson [[hidden email]]
Sent: Tuesday, September 5, 2017 1:07 PM
To: [hidden email]
Subject: Re: Laser-based AF for Nikon Ti-S

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

Thanks Kyle,

We will routinely use dry 20x/40x 0.7-0.9NA objectives.

Darren

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Kyle Michael Douglass
Sent: 05 September 2017 11:20
To: [hidden email]
Subject: Re: Laser-based AF for Nikon Ti-S

*****
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.
*****

My apologies: I meant to say that the objective NA should be higher than the refractive index of the mounting medium so that the laser beam may be totally internally reflected from the coverslip/medium interface.

Kyle

Dr. Kyle M. Douglass
Post-doctoral Researcher
EPFL - The Laboratory of Experimental Biophysics http://leb.epfl.ch/ http://kmdouglass.github.io

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Kyle Michael Douglass [[hidden email]]
Sent: Tuesday, September 5, 2017 12:14 PM
To: [hidden email]
Subject: Re: Laser-based AF for Nikon Ti-S

*****
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 Darren,

I don't know the specifics of the Ti-S, but if it uses an oil objective with NA higher than your mounting medium, then you could try the pgFocus open hardware system from Karl Bellvé.

http://big.umassmed.edu/wiki/index.php/PgFocus

You will need a stage or objective scanner that accepts a 0 - 10 V analog control signal and to setup the IR reference beam optics yourself.

I use this for focus lock on a 60x oil widefield setup and can maintain focus to within 5 - 10 nm standard deviation. It can sometimes be tricky to scan the stage with the focus locked, but otherwise it works well.

Cheers,
Kyle


Dr. Kyle M. Douglass
Post-doctoral Researcher
EPFL - The Laboratory of Experimental Biophysics http://leb.epfl.ch/ http://kmdouglass.github.io

________________________________________
From: Confocal Microscopy List [[hidden email]] on behalf of Darren Thomson [[hidden email]]
Sent: Tuesday, September 5, 2017 11:43 AM
To: [hidden email]
Subject: Laser-based AF for Nikon Ti-S

*****
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 Listserv,

We've been told by Nikon that no hardware based AutoFocus system is compatible with our Nikon Ti-S body and that we have to acquire a new microscope. Has anyone had luck, or suggest alternatives? Software based AF won't do for our throughput applications.

Many thanks for your help,

Best,

Darren Thomson
Manchester Fungal Infection Group
University of Manchester