Olympus ZDC 2 vs. Nikon PFS

[mcammer]

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If there is anyone out there who has had problems with the Nikon PFS system drifting out of focus during experiments or problems with multiple field focus fidelity, would you please contact me privately.  We are trying to assess how common this problem is and, more importantly, want to know any tips to counter these issues.

Thank you!

_________________________________________
Michael Cammer, Assistant Research Scientist
Skirball Institute of Biomolecular Medicine
Lab: [phone]  Cell: [phone]

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[Axel Kurt Preuss]

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Dear Michael
What s your email?
Please say Hello to Rudolfo from me...

Cheers
Axel


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Cammer, Michael
Sent: Sunday, 17 July, 2011 8:17 AM
To: [hidden email]
Subject: Re: Olympus ZDC 2 vs. Nikon PFS

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If there is anyone out there who has had problems with the Nikon PFS system drifting out of focus during experiments or problems with multiple field focus fidelity, would you please contact me privately.  We are trying to assess how common this problem is and, more importantly, want to know any tips to counter these issues.

Thank you!

_________________________________________
Michael Cammer, Assistant Research Scientist Skirball Institute of Biomolecular Medicine
Lab: [phone]  Cell: [phone]

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[mcammer]

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Just wanted to give an update.

We did extensive tests with Tetraspeck beads imaged by TIRF and the Nikon PFS with the 100X TIRF objective held the focus for hours under different temperature conditions.  There was lateral drift of up to 10 um when we turned the heat on and off to check for focus locking during temperature shifts, but the focus held.  We were very happy about this.

However, when we switched to cells on lipid substrates in the exact same chamber, the focus did not hold.

So we are somewhat mystified and continuing to run tests.

Regards,

Michael

From: Craig Brideau [mailto:[hidden email]]
Sent: Monday, July 18, 2011 12:34 AM
To: Cammer, Michael
Subject: Re: Olympus ZDC 2 vs. Nikon PFS

We have a Nikon PFS that mostly works.  We learned a few things early on that help make it more reliable.  It needs #1.5 cover slips.  If you use #1 it will overfocus as it is expecting #1.5 thick coverslips.  Dirt can be a big issue; the lens has to be fairly clean.  Sometimes dirt on the coverslip can throw things off as well.  Keeping the sample level is important, if there is any tilt it gets thrown off.  Finding the initial focus to get it to 'lock on' can require some patience occasionally.  You have to manually adjust the focus and keep trying to engage the PFS system.  It will eventually find the surface but it sometimes takes a few tries.  Usually it will get it on the first try though.  The performance is different for different objectives, so try all the PFS compatible objectives you have to see how they work.
I hope these tips help!

Craig


On Sat, Jul 16, 2011 at 6:16 PM, Cammer, Michael <[hidden email]<mailto:[hidden email]>> wrote:
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If there is anyone out there who has had problems with the Nikon PFS system drifting out of focus during experiments or problems with multiple field focus fidelity, would you please contact me privately.  We are trying to assess how common this problem is and, more importantly, want to know any tips to counter these issues.

Thank you!

_________________________________________
Michael Cammer, Assistant Research Scientist
Skirball Institute of Biomolecular Medicine
Lab: [phone]  Cell: [phone]


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[Tim Feinstein-2]

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For what it's worth, our lab has used two PFS-enabled Nikon scopes for daily multi-XY time-lapse imaging for a couple of years now.  We did not compare PFS with any other system as competing focus control schemes were still largely software-based (and thus less appealing) in 2008.  In our experience lateral drift is almost always caused by the imaging chamber shifting in its mount.  One of our scopes has a spring-loaded chamber holder (in a piezo stage insert) and never experiences lateral drift.  The other scope has a different holder and had issues with XY drif until we glued some strips of rubber band to the sides of the chamber holder to prevent slippage.  That worked until the rubber band strips saturated with immersion oil and had to be changed.

We almost always use 25mm coverslips in Attofluor chambers (Invitrogen), or else Mattek dishes with #1.5 glass.  In our experience PFS lock is easy and fast to acquire after a little practice.  The main impediment is normally small bubbles in the immersion oil.  Since using PFS I have become much more conscious of these - some days I will oil, clean and re-oil an objective once or twice to verify that the tiny bubbles are completely gone.  These can be the most frustrating because acquisition and imaging will work fine but PFS will randomly disengage during imaging as the bubble shifts around.  The other major reasons we lose focus are glass tilt, as others have mentioned, and evaporation of the cell medium.   Reasonable temperature shifts do not affect us much.  

Since glass tilt can be hard to notice, we only ask the PFS to make smallish jumps between each XY position.  If necessary we add an 'empty' XY position to break one large jump into two small ones.  The glass moves much faster while scanning for cells that it does during imaging so a problem can (frustratingly) only appear after you set everything up and press 'run'.   A little foresight prevents that.  

Dirt would certainly be a problem, but I am rather OCD about keeping glass clean.  

I hope this helps,


TF

Timothy Feinstein, PhD
Postdoctoral Fellow
Laboratory for GPCR Biology
Dept. of Pharmacology & Chemical Biology
University of Pittsburgh, School of Medicine
BST W1301, 200 Lothrop St.
Pittsburgh, PA  15261

On Jul 21, 2011, at 10:54 AM, Cammer, Michael wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Just wanted to give an update.
>
> We did extensive tests with Tetraspeck beads imaged by TIRF and the Nikon PFS with the 100X TIRF objective held the focus for hours under different temperature conditions.  There was lateral drift of up to 10 um when we turned the heat on and off to check for focus locking during temperature shifts, but the focus held.  We were very happy about this.
>
> However, when we switched to cells on lipid substrates in the exact same chamber, the focus did not hold.
>
> So we are somewhat mystified and continuing to run tests.
>
> Regards,
>
> Michael
>
> From: Craig Brideau [mailto:[hidden email]]
> Sent: Monday, July 18, 2011 12:34 AM
> To: Cammer, Michael
> Subject: Re: Olympus ZDC 2 vs. Nikon PFS
>
> We have a Nikon PFS that mostly works.  We learned a few things early on that help make it more reliable.  It needs #1.5 cover slips.  If you use #1 it will overfocus as it is expecting #1.5 thick coverslips.  Dirt can be a big issue; the lens has to be fairly clean.  Sometimes dirt on the coverslip can throw things off as well.  Keeping the sample level is important, if there is any tilt it gets thrown off.  Finding the initial focus to get it to 'lock on' can require some patience occasionally.  You have to manually adjust the focus and keep trying to engage the PFS system.  It will eventually find the surface but it sometimes takes a few tries.  Usually it will get it on the first try though.  The performance is different for different objectives, so try all the PFS compatible objectives you have to see how they work.
> I hope these tips help!
>
> Craig
>
>
> On Sat, Jul 16, 2011 at 6:16 PM, Cammer, Michael <[hidden email]<mailto:[hidden email]>> wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
> If there is anyone out there who has had problems with the Nikon PFS system drifting out of focus during experiments or problems with multiple field focus fidelity, would you please contact me privately.  We are trying to assess how common this problem is and, more importantly, want to know any tips to counter these issues.
>
> Thank you!
>
> _________________________________________
> Michael Cammer, Assistant Research Scientist
> Skirball Institute of Biomolecular Medicine
> Lab: [phone]  Cell: [phone]
>
>
> </PRE>
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[Tim Feinstein-2]

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Correction - The glass moves much faster while imaging than it does while scanning for cells.  


TF

Timothy Feinstein, PhD
Postdoctoral Fellow
Laboratory for GPCR Biology
Dept. of Pharmacology & Chemical Biology
University of Pittsburgh, School of Medicine
BST W1301, 200 Lothrop St.
Pittsburgh, PA  15261

On Jul 21, 2011, at 10:54 AM, Cammer, Michael wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Just wanted to give an update.
>
> We did extensive tests with Tetraspeck beads imaged by TIRF and the Nikon PFS with the 100X TIRF objective held the focus for hours under different temperature conditions.  There was lateral drift of up to 10 um when we turned the heat on and off to check for focus locking during temperature shifts, but the focus held.  We were very happy about this.
>
> However, when we switched to cells on lipid substrates in the exact same chamber, the focus did not hold.
>
> So we are somewhat mystified and continuing to run tests.
>
> Regards,
>
> Michael
>
> From: Craig Brideau [mailto:[hidden email]]
> Sent: Monday, July 18, 2011 12:34 AM
> To: Cammer, Michael
> Subject: Re: Olympus ZDC 2 vs. Nikon PFS
>
> We have a Nikon PFS that mostly works.  We learned a few things early on that help make it more reliable.  It needs #1.5 cover slips.  If you use #1 it will overfocus as it is expecting #1.5 thick coverslips.  Dirt can be a big issue; the lens has to be fairly clean.  Sometimes dirt on the coverslip can throw things off as well.  Keeping the sample level is important, if there is any tilt it gets thrown off.  Finding the initial focus to get it to 'lock on' can require some patience occasionally.  You have to manually adjust the focus and keep trying to engage the PFS system.  It will eventually find the surface but it sometimes takes a few tries.  Usually it will get it on the first try though.  The performance is different for different objectives, so try all the PFS compatible objectives you have to see how they work.
> I hope these tips help!
>
> Craig
>
>
> On Sat, Jul 16, 2011 at 6:16 PM, Cammer, Michael <[hidden email]<mailto:[hidden email]>> wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
> If there is anyone out there who has had problems with the Nikon PFS system drifting out of focus during experiments or problems with multiple field focus fidelity, would you please contact me privately.  We are trying to assess how common this problem is and, more importantly, want to know any tips to counter these issues.
>
> Thank you!
>
> _________________________________________
> Michael Cammer, Assistant Research Scientist
> Skirball Institute of Biomolecular Medicine
> Lab: [phone]  Cell: [phone]
>
>
> </PRE>
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[Arvydas Matiukas]

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Dear list,
 
I wonder if anybody tried to simultaneously run two 2p systems
off single Chameleon 2p laser.
 
Additional issue is how to deliver 2p beam to the second system
which we want to setup on a separate optical table. Will a fiber
connection work? Please suggest any potential options that would be
significantly cheaper than buying second Chameleon laser ($160k).
 
 
Thanks,


 
 
 
Arvydas Matiukas, Ph.D.
Director of Confocal&Two-Photon Core
Department of Pharmacology
SUNY Upstate Medical University
766 Irving Ave., WH 3167
Syracuse, NY 13210
tel.: [phone]
fax: [phone]
email: [hidden email]

[Andreas Bruckbauer]

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 I think Zeiss and possibly Olympus offer solutions for this, but as i understand both microscopes have to be on the same table. Focussing a pulsed NIR laser (3W) onto a fiber will most likely vaporise the fiber or are there any special fibers for this application?

best wishes

Andreas

 


 

 

-----Original Message-----
From: Arvydas Matiukas <[hidden email]>
To: CONFOCALMICROSCOPY <[hidden email]>
Sent: Thu, 21 Jul 2011 22:25
Subject: Driving two 2p scopes with a single 2p  laser


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Dear list,

 

I wonder if anybody tried to simultaneously run two 2p systems

off single Chameleon 2p laser.

 

Additional issue is how to deliver 2p beam to the second system

which we want to setup on a separate optical table. Will a fiber

connection work? Please suggest any potential options that would be

significantly cheaper than buying second Chameleon laser ($160k).

 

 

Thanks,





 

 

 

Arvydas Matiukas, Ph.D.

Director of Confocal&Two-Photon Core

Department of Pharmacology

SUNY Upstate Medical University

766 Irving Ave., WH 3167

Syracuse, NY 13210

tel.: [phone]

fax: [phone]

email: [hidden email]


 

[Armstrong, Brian]

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We run two Confocal microscopes off of one Chameleon laser. We split the beam with a polarizing beam splitter, the two beams then go to two AOMs (Zeiss) which controls percent power. We operate the Chameleon by USB to its own computer, that way if someone leaves they do not turn off the laser for the other user.  
We have the entire system on one optical table (8' x 5'), I would not try it on two tables. My understanding is that you do not want to run a 2P beam through a fiber, although I have heard of someone doing it recently, perhaps others can comment about doing this.

The systems work well and we seem to have plenty of power out of a Chameleon 210 for both microscopes.

Hope this helps,

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 Arvydas Matiukas
Sent: Thursday, July 21, 2011 2:25 PM
To: [hidden email]
Subject: Driving two 2p scopes with a single 2p laser

*****
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Dear list,
 
I wonder if anybody tried to simultaneously run two 2p systems
off single Chameleon 2p laser.
 
Additional issue is how to deliver 2p beam to the second system
which we want to setup on a separate optical table. Will a fiber
connection work? Please suggest any potential options that would be
significantly cheaper than buying second Chameleon laser ($160k).
 
 
Thanks,


 
 
 
Arvydas Matiukas, Ph.D.
Director of Confocal&Two-Photon Core
Department of Pharmacology
SUNY Upstate Medical University
766 Irving Ave., WH 3167
Syracuse, NY 13210
tel.: [phone]
fax: [phone]
email: [hidden email]


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[Cameron Nowell]

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Hi Michael,

Could it be because you cells are moving a different amount laterally to
your coverslip? PFS (and Olympus ZDC) just find the coverslip again each
time. They assume your sample stays at the same lateral position in
relation to the coverslip. So if you cover slip moves laterally 10um but
your cells move 20um the PFS will only compensate for the 10um drift and
your cells will still be out of focus.


Cheers
Cam


Cameron J. Nowell
Microscopy Manager
Centre for Advanced Microscopy
Ludwig Institute for Cancer Research Melbourne - Parkville Branch
PO Box 2008
Royal Melbourne Hospital
Victoria, 3050
AUSTRALIA
Office: [phone]
Mobile: [phone]
Fax: [phone]
Facility Website



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Cammer, Michael
Sent: Friday, 22 July 2011 12:55 AM
To: [hidden email]
Subject: Re: Olympus ZDC 2 vs. Nikon PFS

*****
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Just wanted to give an update.

We did extensive tests with Tetraspeck beads imaged by TIRF and the
Nikon PFS with the 100X TIRF objective held the focus for hours under
different temperature conditions.  There was lateral drift of up to 10
um when we turned the heat on and off to check for focus locking during
temperature shifts, but the focus held.  We were very happy about this.

However, when we switched to cells on lipid substrates in the exact same
chamber, the focus did not hold.

So we are somewhat mystified and continuing to run tests.

Regards,

Michael

From: Craig Brideau [mailto:[hidden email]]
Sent: Monday, July 18, 2011 12:34 AM
To: Cammer, Michael
Subject: Re: Olympus ZDC 2 vs. Nikon PFS

We have a Nikon PFS that mostly works.  We learned a few things early on
that help make it more reliable.  It needs #1.5 cover slips.  If you use
#1 it will overfocus as it is expecting #1.5 thick coverslips.  Dirt can
be a big issue; the lens has to be fairly clean.  Sometimes dirt on the
coverslip can throw things off as well.  Keeping the sample level is
important, if there is any tilt it gets thrown off.  Finding the initial
focus to get it to 'lock on' can require some patience occasionally.
You have to manually adjust the focus and keep trying to engage the PFS
system.  It will eventually find the surface but it sometimes takes a
few tries.  Usually it will get it on the first try though.  The
performance is different for different objectives, so try all the PFS
compatible objectives you have to see how they work.
I hope these tips help!

Craig


On Sat, Jul 16, 2011 at 6:16 PM, Cammer, Michael
<[hidden email]<mailto:[hidden email]>> wrote:
*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****
If there is anyone out there who has had problems with the Nikon PFS
system drifting out of focus during experiments or problems with
multiple field focus fidelity, would you please contact me privately.
We are trying to assess how common this problem is and, more
importantly, want to know any tips to counter these issues.

Thank you!

_________________________________________
Michael Cammer, Assistant Research Scientist
Skirball Institute of Biomolecular Medicine
Lab: [phone]  Cell: [phone]


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prohibited. If you have received this email in error please notify the
sender by return email and delete the original message. Please note, the
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The views expressed in this communication are those of the sender and do not necessarily reflect the views of the Ludwig Institute for Cancer Research Ltd.

[Alberto Diaspro]

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Hi
We run two confocal microscopes Leica SP5 - inverted and upright - using a Chameleon at NBT of the Italian Institute of Technology.
Cia
Alby

p.s. we can send set up details

On Jul 21, 2011, at 11:36 PM, Armstrong, Brian wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> We run two Confocal microscopes off of one Chameleon laser. We split the beam with a polarizing beam splitter, the two beams then go to two AOMs (Zeiss) which controls percent power. We operate the Chameleon by USB to its own computer, that way if someone leaves they do not turn off the laser for the other user.  
> We have the entire system on one optical table (8' x 5'), I would not try it on two tables. My understanding is that you do not want to run a 2P beam through a fiber, although I have heard of someone doing it recently, perhaps others can comment about doing this.
>
> The systems work well and we seem to have plenty of power out of a Chameleon 210 for both microscopes.
>
> Hope this helps,
>
> 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 Arvydas Matiukas
> Sent: Thursday, July 21, 2011 2:25 PM
> To: [hidden email]
> Subject: Driving two 2p scopes with a single 2p laser
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear list,
>
> I wonder if anybody tried to simultaneously run two 2p systems
> off single Chameleon 2p laser.
>
> Additional issue is how to deliver 2p beam to the second system
> which we want to setup on a separate optical table. Will a fiber
> connection work? Please suggest any potential options that would be
> significantly cheaper than buying second Chameleon laser ($160k).
>
>
> Thanks,
>
>
>
>
>
> Arvydas Matiukas, Ph.D.
> Director of Confocal&Two-Photon Core
> Department of Pharmacology
> SUNY Upstate Medical University
> 766 Irving Ave., WH 3167
> Syracuse, NY 13210
> tel.: [phone]
> fax: [phone]
> email: [hidden email]
>
>
> ---------------------------------------------------------------------
> *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.
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[Johannes Helm]

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Good morning,

I have installed at different places of the same University two systems -
and am now installing a third one - including two MPLSMs each and sharing
a laser for imaging and another laser for chemical activation. It works
fine.

What I did is to send the resp. laser beams through an achromatic
(achromatic from approx. 680nm - approx. 1064nm, the achromaticity is
important!) 1/2 lambda plate mounted in a motor driven rotation stage. A
polarizing beam splitter cube or Foster prism are mounted behind the
lambda/2 plate.

The motor driving the rotation stage for the lambda/2 plate is controlled
via a PC and a dedicated program. I have solutions from Newport and Standa
and both work equally fine.

The namely PC also controls the resp. two lasers (one Chameleon, one
DeepSee, controlling two lasers of the same type from one PC might be a
problem). I have established calibration curves for the position of the
rotation stage indicating the transmitted power as a function of the motor
position.

With this arrangement, the users can adjust for the required laser power
without having to access the hardware on the optical table (which, for
safety reasons, is hidden behind an enclosure from Al plates).

I think it is a very good idea to avoid having the laser control program
and / or lambda/2 position control installed on the same PC(s) controlling
the MPLSMs. A "neutral" third PC helps to avoid a lot of possible trouble.

It is, to my mind, necessary to avoid any optical fiber when operating
MPLSMs using ultrafast lasers with pulses in the few hundreds fsec or
sub-onehundred fsec range. Thus, both setups should be mounted on one
optical table.

Best wishes,

Johannes

--
P. Johannes Helm, M.Sc. PhD
Seniorengineer
CMBN
University of Oslo
Institute of Basic Medical Science
Department of Anatomy
Postboks 1105 - Blindern
NO-0317 Oslo

Voice: [phone]
Fax: [phone]

WWW: folk.uio.no/jhelm

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Dear list,
>
> I wonder if anybody tried to simultaneously run two 2p systems
> off single Chameleon 2p laser.
>
> Additional issue is how to deliver 2p beam to the second system
> which we want to setup on a separate optical table. Will a fiber
> connection work? Please suggest any potential options that would be
> significantly cheaper than buying second Chameleon laser ($160k).
>
>
> Thanks,
>
>
>
>
>
> Arvydas Matiukas, Ph.D.
> Director of Confocal&Two-Photon Core
> Department of Pharmacology
> SUNY Upstate Medical University
> 766 Irving Ave., WH 3167
> Syracuse, NY 13210
> tel.: [phone]
> fax: [phone]
> email: [hidden email]
>

[Craig Brideau]

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

We did the 1/2 waveplate and polarizing beamsplitter cube trick mentioned by
others.  You don't need a motorized mount for the waveplate though; a manual
one works fine and is much cheaper.  The one problem we found, which is why
we eventually got a second laser, is that the users of the two microscopes
inevitably needed to work at different wavelengths.  As a result only one
microscope was running at a time most days.  On the other hand, it did allow
us to have an inverted and an upright fed from the same laser, so for each
experiment we could use the type of scope best suited for the job.

Craig


On Fri, Jul 22, 2011 at 3:28 AM, P. Johannes Helm
<[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Good morning,
>
> I have installed at different places of the same University two systems -
> and am now installing a third one - including two MPLSMs each and sharing
> a laser for imaging and another laser for chemical activation. It works
> fine.
>
> What I did is to send the resp. laser beams through an achromatic
> (achromatic from approx. 680nm - approx. 1064nm, the achromaticity is
> important!) 1/2 lambda plate mounted in a motor driven rotation stage. A
> polarizing beam splitter cube or Foster prism are mounted behind the
> lambda/2 plate.
>
> The motor driving the rotation stage for the lambda/2 plate is controlled
> via a PC and a dedicated program. I have solutions from Newport and Standa
> and both work equally fine.
>
> The namely PC also controls the resp. two lasers (one Chameleon, one
> DeepSee, controlling two lasers of the same type from one PC might be a
> problem). I have established calibration curves for the position of the
> rotation stage indicating the transmitted power as a function of the motor
> position.
>
> With this arrangement, the users can adjust for the required laser power
> without having to access the hardware on the optical table (which, for
> safety reasons, is hidden behind an enclosure from Al plates).
>
> I think it is a very good idea to avoid having the laser control program
> and / or lambda/2 position control installed on the same PC(s) controlling
> the MPLSMs. A "neutral" third PC helps to avoid a lot of possible trouble.
>
> It is, to my mind, necessary to avoid any optical fiber when operating
> MPLSMs using ultrafast lasers with pulses in the few hundreds fsec or
> sub-onehundred fsec range. Thus, both setups should be mounted on one
> optical table.
>
> Best wishes,
>
> Johannes
>
> --
> P. Johannes Helm, M.Sc. PhD
> Seniorengineer
> CMBN
> University of Oslo
> Institute of Basic Medical Science
> Department of Anatomy
> Postboks 1105 - Blindern
> NO-0317 Oslo
>
> Voice:  [phone]
> Fax:    [phone]
>
> WWW:    folk.uio.no/jhelm
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Dear list,
> >
> > I wonder if anybody tried to simultaneously run two 2p systems
> > off single Chameleon 2p laser.
> >
> > Additional issue is how to deliver 2p beam to the second system
> > which we want to setup on a separate optical table. Will a fiber
> > connection work? Please suggest any potential options that would be
> > significantly cheaper than buying second Chameleon laser ($160k).
> >
> >
> > Thanks,
> >
> >
> >
> >
> >
> > Arvydas Matiukas, Ph.D.
> > Director of Confocal&Two-Photon Core
> > Department of Pharmacology
> > SUNY Upstate Medical University
> > 766 Irving Ave., WH 3167
> > Syracuse, NY 13210
> > tel.: [phone]
> > fax: [phone]
> > email: [hidden email]
> >
>

[SL Smith]

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

The second half of this blog post has some references for sending fs pulses from
Ti:Sapph lasers over fibers:
http://labrigger.com/blog/2010/11/07/fiber-optic-termination/

"It is possible to do so, without massive pulse dispersion, using a hollow core
photonic band-gap fiber (ref, ref). It can also be done with a single mode fiber,
but requires more kit (ref)."

[mcammer]

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

Just wanted to send an update in favor of the Nikon PFS.  We're still not sure about the stability with the live chambers, but on fixed material we are able to use the Large Image or Tiling to image many fields while holding the focus.  We focus on the green channel (in TIRF) and put offsets in the Advanced options of the Lambda to adjust for long UV and IR focal shifts.  Also, the focus shift feature is useful when including brightfield and widefield images to automatically focus at a field above the substrate.  And it goes back to the correct focal plane. One mistake we were making:  if you are using an objective with a correction collar for temperature, make sure it is adjusted for really crisp focus.  
_________________________________________
Michael Cammer, Assistant Research Scientist
Skirball Institute of Biomolecular Medicine
Lab: [phone]  Cell: [phone]




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