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SP5 vs fluoview 1000 for FCS modification ?

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Jean-Pierre CLAMME

SP5 vs fluoview 1000 for FCS modification ?

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi,

I'm wondering if onyone has experience with modifying a SP5 or a Fluoview
1000 for FCS mesures ? I work with Olympus stands before and they generally
offer good access to external ports. But what a bout the SP5 ?

Thanks

JP

Csúcs Gábor

Re: SP5 vs fluoview 1000 for FCS modification ?

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Jean-Pierre,

I've do direct experience with an SP5 but we have a functional FCS setup
on an SP2. As far as I know in this respect there is no difference
between the SP2 and the SP5. There is simply an external port on the
confocal head-where you can insert your detector. As far as I know you
can purchase an FCS unit also from Leica. One of the advantages if you
do this, that you can use the APD-s for imaging and their sensitivity is
superior compared to the PMT-s.

Cheers Gabor

--
Gabor Csucs
Light Microscopy Centre, ETH Zurich
Schafmattstrasse 18, HPM F16
CH-8093, Zurich, Switzerland

Web: www.lmc.ethz.ch
Phone: +41 44 633 6221
Fax: +41 44 632 1298
e-mail: [hidden email]

Michael Weber-4

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Jean-Pierre CLAMME

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Jean-Pierre,

on both systems one needs to install the external port modification (X1
/ port 4 option), in order to bring the light out of the scan-head.
Beside that, it's just another discussion about which system offers
better transmission - dichromatic mirrors or AOBS.

Gabor, can you control the APDs for imaging via the Leica software, or
do you have to use an external solution?

Michael

Jean-Pierre CLAMME wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi,
>
> I'm wondering if onyone has experience with modifying a SP5 or a Fluoview
> 1000 for FCS mesures ? I work with Olympus stands before and they generally
> offer good access to external ports. But what a bout the SP5 ?
>
>
> Thanks
>
> JP

Michael Weber-4

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Jean-Pierre CLAMME

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Jean-Pierre,

on both systems one needs to install the external port modification (X1
/ port 4 option), in order to bring the light out of the scan-head.
Beside that, it's just another discussion about which system offers
better transmission - dichromatic mirrors or AOBS.

Gabor, can you control the APDs for imaging via the Leica software, or
do you have to use an external solution? It's pity that Leica doesn't
offer an internal APD-option including "spectral freedom".

Michael

Jean-Pierre CLAMME wrote:

Michael Weber-4

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Michael Weber-4

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Ts, first rejected, then sent... I don't get it. Sorry for the
double-posting.

Michael

Michael Weber wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Jean-Pierre,
>
> on both systems one needs to install the external port modification (X1
> / port 4 option), in order to bring the light out of the scan-head.
> Beside that, it's just another discussion about which system offers
> better transmission - dichromatic mirrors or AOBS.
>
> Gabor, can you control the APDs for imaging via the Leica software, or
> do you have to use an external solution?
>
> Michael
>
>
> Jean-Pierre CLAMME wrote:
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Hi,
>>
>> I'm wondering if onyone has experience with modifying a SP5 or a Fluoview
>> 1000 for FCS mesures ? I work with Olympus stands before and they
>> generally
>> offer good access to external ports. But what a bout the SP5 ?
>>
>> Thanks
>>
>> JP

Csúcs Gábor

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Michael Weber-4

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Dear Michael,

You can control the APDs from the Leica software - you see them like
"normal" PMTs. I agree with you that it would be nice (especially with
the resonant scanner of the SP5) to have them "behind" the spectral
separation.

Cheers Gabor

--
Gabor Csucs
Light Microscopy Centre, ETH Zurich
Schafmattstrasse 18, HPM F16
CH-8093, Zurich, Switzerland

Web: www.lmc.ethz.ch
Phone: +41 44 633 6221
Fax: +41 44 632 1298
e-mail: [hidden email]

Pan Xiaotao

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Jean-Pierre CLAMME

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi Jean-Pierre,

We did modify a FV300 system for FCS measurement, there is detailed
description about modification with technical drawing in our paper
published on Review of Scientific Instrument, here is the link
(http://dx.doi.org/10.1063/1.2740053 ).

It should not be very difficult if you also need to do APD imaging, in
principle, you can bypass one fluorescence channel and connect that in
control box to APD output (maybe signal conversion needed here). The
modified system is not expensive and its performance is pretty good.

Best Regards,
Xiaotao

Jean-Pierre CLAMME wrote:

Jean-Pierre CLAMME

Re: SP5 vs fluoview 1000 for FCS modification ?

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Could you please send me a pdf version of your paper ?

Thanks

Jean-Pierre

Pan Xiaotao wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi Jean-Pierre,
>
> We did modify a FV300 system for FCS measurement, there is detailed
> description about modification with technical drawing in our paper
> published on Review of Scientific Instrument, here is the link
> (http://dx.doi.org/10.1063/1.2740053 ).
>
> It should not be very difficult if you also need to do APD imaging,
> in principle, you can bypass one fluorescence channel and connect that
> in control box to APD output (maybe signal conversion needed here).
> The modified system is not expensive and its performance is pretty good.
>
> Best Regards,
> Xiaotao
>
>
>
> Jean-Pierre CLAMME wrote:
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Hi,
>>
>> I'm wondering if onyone has experience with modifying a SP5 or a
>> Fluoview
>> 1000 for FCS mesures ? I work with Olympus stands before and they
>> generally
>> offer good access to external ports. But what a bout the SP5 ?
>>
>> Thanks
>>
>> JP
>>
>>
>
>

--
Dr. Jean-Pierre CLAMME
Supervisor 2-Photon Imaging Facility
Dept. of Immunology, IMM-1, R310
The Scripps Research Institute
10550 North Torrey Pines Rd.
La Jolla, California 92037

Lab number: (858)784-8184
Fax number:(858)784-9272

Jean-Pierre CLAMME

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Michael Weber-4

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi,

First thank you for the infos.

Second do you need to go back through the scan heads ? Is it possible to
use another side port to extract the light for example to do cross
correlation ? I worked on two home made systems (FCS + imaging and
single molecule) were we would collect the light directly after the
objective and redirect the signal to multiple APDs. On the SP5 we have
here we have external detectors. Could that kind of approach be used ?

Thanks

JP

Michael Weber wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Jean-Pierre,
>
> on both systems one needs to install the external port modification
> (X1 / port 4 option), in order to bring the light out of the
> scan-head. Beside that, it's just another discussion about which
> system offers better transmission - dichromatic mirrors or AOBS.
>
> Gabor, can you control the APDs for imaging via the Leica software, or
> do you have to use an external solution?
>
> Michael
>
>
> Jean-Pierre CLAMME wrote:
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Hi,
>>
>> I'm wondering if onyone has experience with modifying a SP5 or a
>> Fluoview
>> 1000 for FCS mesures ? I work with Olympus stands before and they
>> generally
>> offer good access to external ports. But what a bout the SP5 ?
>>
>> Thanks
>>
>> JP
>
>

Jean-Pierre CLAMME

Re: SP5 vs fluoview 1000 for FCS modification ?

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

When I say external detectors for the SP5 I mean NDDs.

JP

Jean-Pierre CLAMME wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi,
>
> First thank you for the infos.
>
> Second do you need to go back through the scan heads ? Is it possible
> to use another side port to extract the light for example to do cross
> correlation ? I worked on two home made systems (FCS + imaging and
> single molecule) were we would collect the light directly after the
> objective and redirect the signal to multiple APDs. On the SP5 we have
> here we have external detectors. Could that kind of approach be used ?
>
> Thanks
>
> JP
>
> Michael Weber wrote:
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Jean-Pierre,
>>
>> on both systems one needs to install the external port modification
>> (X1 / port 4 option), in order to bring the light out of the
>> scan-head. Beside that, it's just another discussion about which
>> system offers better transmission - dichromatic mirrors or AOBS.
>>
>> Gabor, can you control the APDs for imaging via the Leica software,
>> or do you have to use an external solution?
>>
>> Michael
>>
>>
>> Jean-Pierre CLAMME wrote:
>>> Search the CONFOCAL archive at
>>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>>
>>> Hi,
>>>
>>> I'm wondering if onyone has experience with modifying a SP5 or a
>>> Fluoview
>>> 1000 for FCS mesures ? I work with Olympus stands before and they
>>> generally
>>> offer good access to external ports. But what a bout the SP5 ?
>>> Thanks
>>>
>>> JP
>>
>>
>

Tony Collins-4

Research technician vacancy at MacBiophotonics

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi Folks,
Please circulate the job posting below to anybody you think would be interested. Please apply through the website.
Thanks,
Tony

A Research Assistant is required to support the management of the day-to-day activities of the robotic cell culture and liquid handling systems at the McMaster Biophotonics Facility. The Facility is an integrated suite of state-of-the-art fluorimetry, microscopy, cytometry and high-content screening systems serving McMaster University and Southern Ontario. The successful candidate will work under the direction of the Manager and Director of the McMaster Biophotonics Facility. Duties include: Support of the Facility's automated cell culture and high content screening platforms including ThermoFisher WorkCells, ArrayScan, PE Opera, Nikon Biostation; Responsible for the routine administration of the high content screening system in the Facility; Assist in the organization of workshops at the Facility; Maintenance, calibration, programming and upgrading instruments in the Facility as directed. The incumbent must have demonstrated experience and knowledge in high content screening including skill in individual support to personnel on the use and applications of high content screening is desirable; Ability to organize and conduct imaging workshops, and willingness to work effectively with researchers with diverse goals and experience is essential; Experience in programming for image processing and acquisition routines; Experience in cell culture and molecular biology are desirable. Applicants must possess a minimum BSc degree in biology, chemistry, biochemistry, physics or medical engineering and appropriate experience.

http://www.workingatmcmaster.ca/careers/job_details.php?PHPSESSID=8d63c417c23654624eed50909255c6ba&selected_job=3467

Tony J. Collins, Ph.D.
McMaster Biophotonics Facility
Dept. Biochemistry and Biomedical Sciences HSC 4H21A
McMaster University, Hamilton, ON, Canada, L8N 3Z5
[hidden email] www.macbiophotonics.ca

ian gibbins

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Csúcs Gábor

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

That's right, Gabor.

We have an SP5 with a dual channel FCS unit attached. The version we
have was supplied by Leica and is made by ISS. The Leica software has a
pretty good FCS "wizard" that manages the interface between the Leica
and the ISS gear, which is run off a second computer. After a very
shakey start, it now works well.

I think it would be possible to put your own FCS unit on the external
port and control the lightpath selection, scan parameters manually, but
you'd need to check, I reckon.

IAN

On Wednesday, June 18, 2008, at 03:18 PM, Csucs Gabor wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Dear Jean-Pierre,
>
> I've do direct experience with an SP5 but we have a functional FCS
> setup on an SP2. As far as I know in this respect there is no
> difference between the SP2 and the SP5. There is simply an external
> port on the confocal head-where you can insert your detector. As far
> as I know you can purchase an FCS unit also from Leica. One of the
> advantages if you do this, that you can use the APD-s for imaging and
> their sensitivity is superior compared to the PMT-s.
>
> Cheers Gabor
>
> --
> Gabor Csucs Light Microscopy Centre, ETH Zurich
> Schafmattstrasse 18, HPM F16 CH-8093, Zurich, Switzerland
>
> Web: www.lmc.ethz.ch
> Phone: +41 44 633 6221
> Fax: +41 44 632 1298
> e-mail: [hidden email]
>
>

* * * * * * * * * * *
Prof Ian Gibbins
Anatomy & Histology
Flinders University
GPO Box 2100
Adelaide SA 5001
AUSTRALIA

[hidden email]
voice: +61-8-8204 5271
fax: +61-8-8277 0085

http://som.flinders.edu.au/FUSA/Anatomy/
http://www.flinders.edu.au/neuroscience

ian gibbins

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Jean-Pierre CLAMME

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

The light path on the SP5 goes to the side port via the scan mirrors
but not the spectral separation path. Going via the scan mirrors allows
you to place the beam at a series of locations when collecting a set of
FCS data (though you could do this by moving the stage with a simply
parked beam I suppose). The set-up we have uses a dichroic and barrier
filters to separate emission bands to two APDs. It works well, but, of
course, you have to change the filter / mirror combinations if you
change your cross-correlation fluorophores, but that is pretty easy too.

IAN

On Thursday, June 19, 2008, at 02:45 AM, Jean-Pierre CLAMME wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> Hi,
>
> First thank you for the infos.
>
> Second do you need to go back through the scan heads ? Is it possible
> to use another side port to extract the light for example to do cross
> correlation ? I worked on two home made systems (FCS + imaging and
> single molecule) were we would collect the light directly after the
> objective and redirect the signal to multiple APDs. On the SP5 we have
> here we have external detectors. Could that kind of approach be used > ?
>
> Thanks
>
> JP
>
> Michael Weber wrote:
>> Search the CONFOCAL archive at
>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>
>> Jean-Pierre,
>>
>> on both systems one needs to install the external port modification
>> (X1 / port 4 option), in order to bring the light out of the
>> scan-head. Beside that, it's just another discussion about which
>> system offers better transmission - dichromatic mirrors or AOBS.
>>
>> Gabor, can you control the APDs for imaging via the Leica software,
>> or do you have to use an external solution?
>>
>> Michael
>>
>>
>> Jean-Pierre CLAMME wrote:
>>> Search the CONFOCAL archive at
>>> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>>>
>>> Hi,
>>>
>>> I'm wondering if onyone has experience with modifying a SP5 or a
>>> Fluoview
>>> 1000 for FCS mesures ? I work with Olympus stands before and they
>>> generally
>>> offer good access to external ports. But what a bout the SP5 ?
>>> Thanks
>>>
>>> JP
>>
>>
>
> --
> Dr. Jean-Pierre CLAMME
> Supervisor 2-Photon Imaging Facility Dept. of Immunology, IMM-1, R310
> The Scripps Research Institute
> 10550 North Torrey Pines Rd.
> La Jolla, California 92037
>
> Lab number: (858)784-8184
> Fax number:(858)784-9272
>
>

Jean-Pierre CLAMME

Re: SP5 vs fluoview 1000 for FCS modification ?

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

This is what we were doing on our system. We would choose a point and
then move the table to bring that point under the parked beam.
I'm just wondering, is the fact that scanning moves the beam changing
the focal volume from one point to the other ? Or is it negligible ?

Thanks

JP
> The light path on the SP5 goes to the side port via the scan mirrors
> but not the spectral separation path. Going via the scan mirrors
> allows you to place the beam at a series of locations when collecting
> a set of FCS data (though you could do this by moving the stage with a
> simply parked beam I suppose).
This is what we were doing on our system. We would choose a point and
then move the table to bring that point under the parked beam.
I'm just wondering Is the fact that when you use the scanner, you but
the fact that when you use the scanner,

>
> [hidden email]
> voice: +61-8-8204 5271
> fax: +61-8-8277 0085
>
> http://som.flinders.edu.au/FUSA/Anatomy/
> http://www.flinders.edu.au/neuroscience
>

ian gibbins

Re: SP5 vs fluoview 1000 for FCS modification ?

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

We've measured that and it's negligible under our usual conditions,
which is a relief...

IAN

On Thursday, June 19, 2008, at 08:54 AM, Jean-Pierre CLAMME wrote:

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> This is what we were doing on our system. We would choose a point and
> then move the table to bring that point under the parked beam.
> I'm just wondering, is the fact that scanning moves the beam
> changing the focal volume from one point to the other ? Or is it
> negligible ?
>
> Thanks
>
> JP
>> The light path on the SP5 goes to the side port via the scan mirrors
>> but not the spectral separation path. Going via the scan mirrors
>> allows you to place the beam at a series of locations when collecting
>> a set of FCS data (though you could do this by moving the stage with
>> a simply parked beam I suppose).
> This is what we were doing on our system. We would choose a point and
> then move the table to bring that point under the parked beam.
> I'm just wondering Is the fact that when you use the scanner, you but
> the fact that when you use the scanner,
>
>
>>
>> [hidden email]
>> voice: +61-8-8204 5271
>> fax: +61-8-8277 0085
>>
>> http://som.flinders.edu.au/FUSA/Anatomy/
>> http://www.flinders.edu.au/neuroscience
>>
>
>

Sudipta Maiti

Re: SP5 vs fluoview 1000 for FCS modification ?

In reply to this post by Jean-Pierre CLAMME

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

There is also a way to use external detectors, pretty much with any confocal
microscope, without worrying about alignment issues. We have used an
external APD to perform FCS at any chosen position in the image. More
details are in "Fluorescence correlation microscopy with real-time
alignment readout", Kaushalya et al., Applied Optics, vol. 44 (2005) p.3262.
You may contact me off the list if you need to know more.
Sudipta

On Wed, 18 Jun 2008 16:24:49 -0700, Jean-Pierre CLAMME wrote

> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> This is what we were doing on our system. We would choose a point
> and then move the table to bring that point under the parked beam.
> I'm just wondering, is the fact that scanning moves the beam
> changing the focal volume from one point to the other ? Or is it
> negligible ?
>
> Thanks
>
> JP
> > The light path on the SP5 goes to the side port via the scan mirrors
> > but not the spectral separation path. Going via the scan mirrors
> > allows you to place the beam at a series of locations when collecting
> > a set of FCS data (though you could do this by moving the stage with a
> > simply parked beam I suppose).
> This is what we were doing on our system. We would choose a point
> and then move the table to bring that point under the parked beam.
> I'm just wondering Is the fact that when you use the scanner, you
> but the fact that when you use the scanner,
>
> >
> > [hidden email]
> > voice: +61-8-8204 5271
> > fax: +61-8-8277 0085
> >
> > http://som.flinders.edu.au/FUSA/Anatomy/
> > http://www.flinders.edu.au/neuroscience
> >

Dr. Sudipta Maiti
Associate Professor
Dept. of Chemical Sciences
Tata Institute of Fundamental Research
Homi Bhabha Raod, Colaba, Mumbai 400005
Ph. 91-22-2278-2716 / 2539
Fax: 91-22-2280-4610
alternate e-mail: [hidden email]
url: www.tifr.res.in/~biophotonics