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405 nm laser via NIR port on Zeiss 510

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Phillips, Thomas E.

405 nm laser via NIR port on Zeiss 510

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The NIR laser for two photon excitation on our Zeiss 510 died on us. The repair would be at least 60K. We are trying to find funds for a new confocal so I don't know if it is worth repairing. The biggest practical problem is that we can't do DAPI excitation on our routine preps. Has anyone tried retrofitting a 405 nm laser through the NIR port? Are there optical components/mirrors in the pathway that only work with longer wavelengths? Any thoughts on this are welcome. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
[hidden email]<mailto:[hidden email]>

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

Craig Brideau

Re: 405 nm laser via NIR port on Zeiss 510

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For starters, what do you have for a Dichroic that could pipe in the 405 and
let the rest of the visible pass? Most 2p dichroics split off at around 680
or 700nm to allow the IR to pass and the emission light to reflect, or
vice-versa.

Craig

On Mon, May 9, 2011 at 1:16 PM, Phillips, Thomas E.
<[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> The NIR laser for two photon excitation on our Zeiss 510 died on us. The
> repair would be at least 60K. We are trying to find funds for a new confocal
> so I don't know if it is worth repairing. The biggest practical problem is
> that we can't do DAPI excitation on our routine preps. Has anyone tried
> retrofitting a 405 nm laser through the NIR port? Are there optical
> components/mirrors in the pathway that only work with longer wavelengths?
> Any thoughts on this are welcome. Tom
>
>
> Thomas E. Phillips, Ph.D
> Professor of Biological Sciences
> Director, Molecular Cytology Core
> 2 Tucker Hall
> University of Missouri
> Columbia, MO 65211-7400
> 573-882-4712 (office)
> 573-882-0123 (fax)
> [hidden email]<mailto:[hidden email]>
>
> http://www.biology.missouri.edu/faculty/phillips.html
> http://www.biotech.missouri.edu/mcc/
>

Armstrong, Brian

Re: 405 nm laser via NIR port on Zeiss 510

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

Craig makes a good point. The HFT700/488 or KP650 (for example) that you have been using for your 2P laser will not work for a 405 line. Therefore you would need to change the dichroics. This is not done in the field and you would need to send your scan-head to the factory in Germany. You would also need to think about how to get your laser into the fiber (AOTF?), how to adjust the power (AOM is no longer useful), and whether or not you have the secondary dichroics that will pass the emitted light.

You should talk to Zeiss before considering this idea any further.

Cheers,

Brian D Armstrong PhD
Assistant Research Professor
Light Microscopy Core
Beckman Research Institute
City of Hope
Dept of Neuroscience
1450 E Duarte Rd
Duarte, CA 91010
626-256-4673 x62872

http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Craig Brideau
Sent: Monday, May 09, 2011 2:52 PM
To: [hidden email]
Subject: Re: 405 nm laser via NIR port on Zeiss 510

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

For starters, what do you have for a Dichroic that could pipe in the 405 and
let the rest of the visible pass? Most 2p dichroics split off at around 680
or 700nm to allow the IR to pass and the emission light to reflect, or
vice-versa.

Craig

On Mon, May 9, 2011 at 1:16 PM, Phillips, Thomas E.
<[hidden email]>wrote:

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Craig Brideau

Re: 405 nm laser via NIR port on Zeiss 510

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

The primary dichroic is typically the largest problem. Our Nikon technician
went through some fairly heroic efforts to install a dichroic into an A1
FN1. We had the filter custom made at significant expense for our
ultrabroadband Ti:Saph laser (Octavius), so the possibility of damaging the
filter during installation was a critical concern. After the technician
partly disassembled the scan head in our lab, we ended up drafting a nice
lady with very tiny fingers who did needlepoint and crocheting as a hobby to
finally get the filter into place in the A1 scan head. This was something
normally done at the factory... That said, it did work out for us and Nikon
was very accommodating in helping us with the modification.
Regarding piping the laser in to the Zeiss scope: I was assuming your laser
could be brought in free space. If you have to go through a fiber then you
probably won't be able to do it due to dispersion in the glass fiber. If
you have a prechirping unit then you *might* be able to pull it off, but it
will be tricky.

Craig

On Mon, May 9, 2011 at 4:06 PM, Armstrong, Brian <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Craig makes a good point. The HFT700/488 or KP650 (for example) that you
> have been using for your 2P laser will not work for a 405 line. Therefore
> you would need to change the dichroics. This is not done in the field and
> you would need to send your scan-head to the factory in Germany. You would
> also need to think about how to get your laser into the fiber (AOTF?), how
> to adjust the power (AOM is no longer useful), and whether or not you have
> the secondary dichroics that will pass the emitted light.
>
> You should talk to Zeiss before considering this idea any further.
>
> Cheers,
>
>
> Brian D Armstrong PhD
> Assistant Research Professor
> Light Microscopy Core
> Beckman Research Institute
> City of Hope
> Dept of Neuroscience
> 1450 E Duarte Rd
> Duarte, CA 91010
> 626-256-4673 x62872
>
>
> http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Craig Brideau
> Sent: Monday, May 09, 2011 2:52 PM
> To: [hidden email]
> Subject: Re: 405 nm laser via NIR port on Zeiss 510
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> For starters, what do you have for a Dichroic that could pipe in the 405
> and
> let the rest of the visible pass? Most 2p dichroics split off at around
> 680
> or 700nm to allow the IR to pass and the emission light to reflect, or
> vice-versa.
>
> Craig
>
>
> On Mon, May 9, 2011 at 1:16 PM, Phillips, Thomas E.
> <[hidden email]>wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > The NIR laser for two photon excitation on our Zeiss 510 died on us. The
> > repair would be at least 60K. We are trying to find funds for a new
> confocal
> > so I don't know if it is worth repairing. The biggest practical problem
> is
> > that we can't do DAPI excitation on our routine preps. Has anyone tried
> > retrofitting a 405 nm laser through the NIR port? Are there optical
> > components/mirrors in the pathway that only work with longer wavelengths?
> > Any thoughts on this are welcome. Tom
> >
> >
> > Thomas E. Phillips, Ph.D
> > Professor of Biological Sciences
> > Director, Molecular Cytology Core
> > 2 Tucker Hall
> > University of Missouri
> > Columbia, MO 65211-7400
> > 573-882-4712 (office)
> > 573-882-0123 (fax)
> > [hidden email]<mailto:[hidden email]>
> >
> > http://www.biology.missouri.edu/faculty/phillips.html
> > http://www.biotech.missouri.edu/mcc/
> >
>
>
> ---------------------------------------------------------------------
> SECURITY/CONFIDENTIALITY WARNING:
> This message and any attachments are intended solely for the individual or
> entity to which they are addressed. This communication may contain
> information that is privileged, confidential, or exempt from disclosure
> under applicable law (e.g., personal health information, research data,
> financial information). Because this e-mail has been sent without
> encryption, individuals other than the intended recipient may be able to
> view the information, forward it to others or tamper with the information
> without the knowledge or consent of the sender. If you are not the intended
> recipient, or the employee or person responsible for delivering the message
> to the intended recipient, any dissemination, distribution or copying of the
> communication is strictly prohibited. If you received the communication in
> error, please notify the sender immediately by replying to this message and
> deleting the message and any accompanying files from your system. If, due to
> the security risks, you do not wish to receive further communications via
> e-mail, please reply to this message and inform the sender that you do not
> wish to receive further e-mail from the sender.
>
> ---------------------------------------------------------------------
>

Craig Brideau

Re: 405 nm laser via NIR port on Zeiss 510

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

Whoops! Got mixed up regarding what laser needed to be brought in: the 405
can be piped in via fiber but it will be fairly important that the fiber is
single-mode at 405nm.

Craig

On Mon, May 9, 2011 at 5:29 PM, Craig Brideau <[hidden email]>wrote:

> The primary dichroic is typically the largest problem. Our Nikon
> technician went through some fairly heroic efforts to install a dichroic
> into an A1 FN1. We had the filter custom made at significant expense for
> our ultrabroadband Ti:Saph laser (Octavius), so the possibility of damaging
> the filter during installation was a critical concern. After the technician
> partly disassembled the scan head in our lab, we ended up drafting a nice
> lady with very tiny fingers who did needlepoint and crocheting as a hobby to
> finally get the filter into place in the A1 scan head. This was something
> normally done at the factory... That said, it did work out for us and Nikon
> was very accommodating in helping us with the modification.
> Regarding piping the laser in to the Zeiss scope: I was assuming your laser
> could be brought in free space. If you have to go through a fiber then you
> probably won't be able to do it due to dispersion in the glass fiber. If
> you have a prechirping unit then you *might* be able to pull it off, but it
> will be tricky.
>
> Craig
>
>
>
> On Mon, May 9, 2011 at 4:06 PM, Armstrong, Brian <[hidden email]>wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Craig makes a good point. The HFT700/488 or KP650 (for example) that you
>> have been using for your 2P laser will not work for a 405 line. Therefore
>> you would need to change the dichroics. This is not done in the field and
>> you would need to send your scan-head to the factory in Germany. You would
>> also need to think about how to get your laser into the fiber (AOTF?), how
>> to adjust the power (AOM is no longer useful), and whether or not you have
>> the secondary dichroics that will pass the emitted light.
>>
>> You should talk to Zeiss before considering this idea any further.
>>
>> Cheers,
>>
>>
>> Brian D Armstrong PhD
>> Assistant Research Professor
>> Light Microscopy Core
>> Beckman Research Institute
>> City of Hope
>> Dept of Neuroscience
>> 1450 E Duarte Rd
>> Duarte, CA 91010
>> 626-256-4673 x62872
>>
>>
>> http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List [mailto:[hidden email]]
>> On Behalf Of Craig Brideau
>> Sent: Monday, May 09, 2011 2:52 PM
>> To: [hidden email]
>> Subject: Re: 405 nm laser via NIR port on Zeiss 510
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> For starters, what do you have for a Dichroic that could pipe in the 405
>> and
>> let the rest of the visible pass? Most 2p dichroics split off at around
>> 680
>> or 700nm to allow the IR to pass and the emission light to reflect, or
>> vice-versa.
>>
>> Craig
>>
>>
>> On Mon, May 9, 2011 at 1:16 PM, Phillips, Thomas E.
>> <[hidden email]>wrote:
>>
>> > *****
>> > To join, leave or search the confocal microscopy listserv, go to:
>> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> > *****
>> >
>> > The NIR laser for two photon excitation on our Zeiss 510 died on us. The
>> > repair would be at least 60K. We are trying to find funds for a new
>> confocal
>> > so I don't know if it is worth repairing. The biggest practical problem
>> is
>> > that we can't do DAPI excitation on our routine preps. Has anyone tried
>> > retrofitting a 405 nm laser through the NIR port? Are there optical
>> > components/mirrors in the pathway that only work with longer
>> wavelengths?
>> > Any thoughts on this are welcome. Tom
>> >
>> >
>> > Thomas E. Phillips, Ph.D
>> > Professor of Biological Sciences
>> > Director, Molecular Cytology Core
>> > 2 Tucker Hall
>> > University of Missouri
>> > Columbia, MO 65211-7400
>> > 573-882-4712 (office)
>> > 573-882-0123 (fax)
>> > [hidden email]<mailto:[hidden email]>
>> >
>> > http://www.biology.missouri.edu/faculty/phillips.html
>> > http://www.biotech.missouri.edu/mcc/
>> >
>>
>>
>> ---------------------------------------------------------------------
>> SECURITY/CONFIDENTIALITY WARNING:
>> This message and any attachments are intended solely for the individual or
>> entity to which they are addressed. This communication may contain
>> information that is privileged, confidential, or exempt from disclosure
>> under applicable law (e.g., personal health information, research data,
>> financial information). Because this e-mail has been sent without
>> encryption, individuals other than the intended recipient may be able to
>> view the information, forward it to others or tamper with the information
>> without the knowledge or consent of the sender. If you are not the intended
>> recipient, or the employee or person responsible for delivering the message
>> to the intended recipient, any dissemination, distribution or copying of the
>> communication is strictly prohibited. If you received the communication in
>> error, please notify the sender immediately by replying to this message and
>> deleting the message and any accompanying files from your system. If, due to
>> the security risks, you do not wish to receive further communications via
>> e-mail, please reply to this message and inform the sender that you do not
>> wish to receive further e-mail from the sender.
>>
>> ---------------------------------------------------------------------
>>
>
>

Axel Kurt Preuss

Re: 405 nm laser via NIR port on Zeiss 510

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

We do "DAPI piping" via standard HBO (!) . Drawback: Pinhole has to be at max to collect enough signal, so it s not so much confocal
Advantage: Cheap (zero costs), efficient, good enough for recognizing nuclei , and it can be done in seconds without any installations
Cheers
Axel

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Craig Brideau
Sent: Tuesday, 10 May, 2011 7:31 AM
To: [hidden email]
Subject: Re: 405 nm laser via NIR port on Zeiss 510

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

Whoops! Got mixed up regarding what laser needed to be brought in: the 405 can be piped in via fiber but it will be fairly important that the fiber is single-mode at 405nm.

Craig

On Mon, May 9, 2011 at 5:29 PM, Craig Brideau <[hidden email]>wrote:

> The primary dichroic is typically the largest problem. Our Nikon
> technician went through some fairly heroic efforts to install a
> dichroic into an A1 FN1. We had the filter custom made at significant
> expense for our ultrabroadband Ti:Saph laser (Octavius), so the
> possibility of damaging the filter during installation was a critical
> concern. After the technician partly disassembled the scan head in
> our lab, we ended up drafting a nice lady with very tiny fingers who
> did needlepoint and crocheting as a hobby to finally get the filter
> into place in the A1 scan head. This was something normally done at
> the factory... That said, it did work out for us and Nikon was very accommodating in helping us with the modification.
> Regarding piping the laser in to the Zeiss scope: I was assuming your
> laser could be brought in free space. If you have to go through a
> fiber then you probably won't be able to do it due to dispersion in
> the glass fiber. If you have a prechirping unit then you *might* be
> able to pull it off, but it will be tricky.
>
> Craig
>
>
>
> On Mon, May 9, 2011 at 4:06 PM, Armstrong, Brian <[hidden email]>wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Craig makes a good point. The HFT700/488 or KP650 (for example) that
>> you have been using for your 2P laser will not work for a 405 line.
>> Therefore you would need to change the dichroics. This is not done in
>> the field and you would need to send your scan-head to the factory in
>> Germany. You would also need to think about how to get your laser
>> into the fiber (AOTF?), how to adjust the power (AOM is no longer
>> useful), and whether or not you have the secondary dichroics that will pass the emitted light.
>>
>> You should talk to Zeiss before considering this idea any further.
>>
>> Cheers,
>>
>>
>> Brian D Armstrong PhD
>> Assistant Research Professor
>> Light Microscopy Core
>> Beckman Research Institute
>> City of Hope
>> Dept of Neuroscience
>> 1450 E Duarte Rd
>> Duarte, CA 91010
>> 626-256-4673 x62872
>>
>>
>> http://www.cityofhope.org/research/support/Light-Microscopy-Digital-I
>> maging/Pages/default.aspx
>>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List
>> [mailto:[hidden email]]
>> On Behalf Of Craig Brideau
>> Sent: Monday, May 09, 2011 2:52 PM
>> To: [hidden email]
>> Subject: Re: 405 nm laser via NIR port on Zeiss 510
>>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> For starters, what do you have for a Dichroic that could pipe in the
>> 405 and let the rest of the visible pass? Most 2p dichroics split
>> off at around
>> 680
>> or 700nm to allow the IR to pass and the emission light to reflect,
>> or vice-versa.
>>
>> Craig
>>
>>
>> On Mon, May 9, 2011 at 1:16 PM, Phillips, Thomas E.
>> <[hidden email]>wrote:
>>
>> > *****
>> > To join, leave or search the confocal microscopy listserv, go to:
>> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> > *****
>> >
>> > The NIR laser for two photon excitation on our Zeiss 510 died on
>> > us. The repair would be at least 60K. We are trying to find funds
>> > for a new
>> confocal
>> > so I don't know if it is worth repairing. The biggest practical
>> > problem
>> is
>> > that we can't do DAPI excitation on our routine preps. Has anyone
>> > tried retrofitting a 405 nm laser through the NIR port? Are there
>> > optical components/mirrors in the pathway that only work with
>> > longer
>> wavelengths?
>> > Any thoughts on this are welcome. Tom
>> >
>> >
>> > Thomas E. Phillips, Ph.D
>> > Professor of Biological Sciences
>> > Director, Molecular Cytology Core
>> > 2 Tucker Hall
>> > University of Missouri
>> > Columbia, MO 65211-7400
>> > 573-882-4712 (office)
>> > 573-882-0123 (fax)
>> > [hidden email]<mailto:[hidden email]>
>> >
>> > http://www.biology.missouri.edu/faculty/phillips.html
>> > http://www.biotech.missouri.edu/mcc/
>> >
>>
>>
>> ---------------------------------------------------------------------
>> SECURITY/CONFIDENTIALITY WARNING:
>> This message and any attachments are intended solely for the
>> individual or entity to which they are addressed. This communication
>> may contain information that is privileged, confidential, or exempt
>> from disclosure under applicable law (e.g., personal health
>> information, research data, financial information). Because this
>> e-mail has been sent without encryption, individuals other than the
>> intended recipient may be able to view the information, forward it to
>> others or tamper with the information without the knowledge or
>> consent of the sender. If you are not the intended recipient, or the
>> employee or person responsible for delivering the message to the
>> intended recipient, any dissemination, distribution or copying of the
>> communication is strictly prohibited. If you received the
>> communication in error, please notify the sender immediately by
>> replying to this message and deleting the message and any
>> accompanying files from your system. If, due to the security risks,
>> you do not wish to receive further communications via e-mail, please reply to this message and inform the sender that you do not wish to receive further e-mail from the sender.
>>
>> ---------------------------------------------------------------------
>>
>
>

Note: This message may contain confidential information. If this Email/Fax has been sent to you by mistake, please notify the sender and delete it immediately. Thank you.

Straatman, Kees (Dr.)

ImageJ workshops 21-22 July

In reply to this post by Craig Brideau

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

Dear list members,

ImageJ is a public domain image processing and analysis program written in Java. It is freely available and used by many scientists all over the world. I will organize two one day ImageJ workshops 'Introduction to ImageJ' on 21 July and 'Writing macros in ImageJ' on 22 July 2011 at the University in Leicester, UK.

The workshops are focussed on scientists who want to use or have started to use ImageJ but are not sure how to make best use of it.

Please visit http://www.le.ac.uk/biochem/microscopy/ImageJ2011.html for further information.

Best wishes

Kees

Dr Ir K.R. Straatman
Senior Experimental Officer
Centre for Core Biotechnology Services
College of Medicine, Biological Sciences and Psychology
http://www.le.ac.uk/biochem/microscopy/home.html