Re: Recommendations for commercial multi-photon system purchase

[Bogdan Stoica]

Hi fellows,

I am also in the last stages of choosing a confocal/multiphoton system for
our laboratory. After seeing multiple demos i am thorn between an Olympus
FV1000MPE system and an Leica SP5 II. Our needs are to be able to do both
confocal images on fixed and live in vitro cells and live in vivo rodent
brain imaging. Both systems come with a UV, visible Argon four lines (488
etc), laser diode 560 and 633 and IR laser (Chameleon for Leica and Maitai
for Olympus), a complete set of Apo objectives as well as the famous 25x NA
1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes are
inverted and the system also includes 2 NDD for Leica and 4 NDD in the case
for Olympus.
Fro what I could see the Leica software appeared more user friendly and I
would give their system a better feel for confocal imaging and manufacturers
potential upgrades- the system I am pricing does not include the AOBS
feature, "white laser" or resonant scanner but those could be added later.
On the other hand the Olympus seems a better choice for multiphoton imaging
and an easier system to tinker with. The tech support and managers for both
companies seemed outstanding to me here in the Washington DC-Baltimore area.
I found interesting that I was not able to find any info on "how much" such
a system should cost and after intense "negotiations" the quites range on my
case around the ~700k mark with Olympus a little bellow and Leica a little
above.
I was wondering if people who recently had to make a similar decision can
offer me some guidance both on the merits of their systems once they were
installed as well as on the price, support etc.
Thank you very much,

Bogdan

[Cameron, Lisa]

Bogdan,

Are you considering pre-churp on the IR laser?

Did you actually compare the Olympus 25x objective and the Leica 20x?
I have not compared them myself, but have heard raves about the Olympus 25x.

How many NDD channels will you need? the difference in 2 NDDs vs. 4 NDDs should
be significant in price, so for the 2 systems you mentioned, I would say the
Olympus is a better price. When you are comparing pricing, you need to compare
equivalent features/parts. Also if you are leaning at all toward the Leica, I
would seriously consider an AOBS - this is the technology that separates them
from the others.

I do not have personal experience with in vivo rodent brain imaging, but it is
my understanding that an upright microscope would be the recommended
configuration. But, if you need to do confocal imaging on live cells, I guess an
inverted would be the best.

I recently went through a similar process of evaluation of systems and would be
happy to compare notes offline or over the phone.

- Lisa

[hidden email]



-----Original Message-----
From: Confocal Microscopy List on behalf of Bogdan Stoica
Sent: Sun 8/23/2009 12:11 PM
To: [hidden email]
Subject: Re: [CONFOCALMICROSCOPY] Recommendations for commercial multi-photon
system purchase
 
Hi fellows,

I am also in the last stages of choosing a confocal/multiphoton system for
our laboratory. After seeing multiple demos i am thorn between an Olympus
FV1000MPE system and an Leica SP5 II. Our needs are to be able to do both
confocal images on fixed and live in vitro cells and live in vivo rodent
brain imaging. Both systems come with a UV, visible Argon four lines (488
etc), laser diode 560 and 633 and IR laser (Chameleon for Leica and Maitai
for Olympus), a complete set of Apo objectives as well as the famous 25x NA
1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes are
inverted and the system also includes 2 NDD for Leica and 4 NDD in the case
for Olympus.
Fro what I could see the Leica software appeared more user friendly and I
would give their system a better feel for confocal imaging and manufacturers
potential upgrades- the system I am pricing does not include the AOBS
feature, "white laser" or resonant scanner but those could be added later.
On the other hand the Olympus seems a better choice for multiphoton imaging
and an easier system to tinker with. The tech support and managers for both
companies seemed outstanding to me here in the Washington DC-Baltimore area.
I found interesting that I was not able to find any info on "how much" such
a system should cost and after intense "negotiations" the quites range on my
case around the ~700k mark with Olympus a little bellow and Leica a little
above.
I was wondering if people who recently had to make a similar decision can
offer me some guidance both on the merits of their systems once they were
installed as well as on the price, support etc.
Thank you very much,

Bogdan



The information in this e-mail is intended only for the person to whom it is
addressed. If you believe this e-mail was sent to you in error and the e-mail
contains patient information, please contact the Partners Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you in error
but does not contain patient information, please contact the sender and properly
dispose of the e-mail.

[Bogdan Stoica]

Dear Lisa,

Thank you for your kind response.
 Let me adress the issue you raised in turn:
-I did not get the chance to compare side by side the Olympus 25x vs. Leica
20x. I could only met with people that had rave reviews about the 25x so I
would tend to five Olympus a slight advantage in this regard.
-initially both systems had 4 NDDs but Leica had to leave only 2 to meet our
budget. I saw people that having 4 NDDs coupled to 2 IR lasers and I was
told me that even with one laser as in our case more NDDs could be
beneficial for recording other channels such as intrinsic fluorescence etc.
Again an extra point for Olympus.
-You are right to point that for Leica features such as the AOBS as well as
white laser should be unique advantages. The problem is that they would put
the system outside our immediate budget although the fact that these
upgrades are available in the future including as on-side upgrades for the
AOBS justify an extra advantage for Leica.
-Indeed, the need to do live cell imaging is directing us toward an inverted
system.
-the IR laser are the Maitai deepsee for olympus and the Chameleon Ultra II
for the Leica.

Bogdan

[Bogdan Stoica]

Dear Lisa,

I answered to your post online but I would really appreciate to speak
with you directly. Please let me know if there is a good time to call
you tomorrow and if so what phone number to use.

Thank you,

Bogdan

Cameron, Lisa wrote:

> Bogdan,
>
> Are you considering pre-churp on the IR laser?
>
> Did you actually compare the Olympus 25x objective and the Leica 20x?
> I have not compared them myself, but have heard raves about the Olympus 25x.
>
> How many NDD channels will you need? the difference in 2 NDDs vs. 4 NDDs should
> be significant in price, so for the 2 systems you mentioned, I would say the
> Olympus is a better price. When you are comparing pricing, you need to compare
> equivalent features/parts. Also if you are leaning at all toward the Leica, I
> would seriously consider an AOBS - this is the technology that separates them
> from the others.
>
> I do not have personal experience with in vivo rodent brain imaging, but it is
> my understanding that an upright microscope would be the recommended
> configuration. But, if you need to do confocal imaging on live cells, I guess an
> inverted would be the best.
>
> I recently went through a similar process of evaluation of systems and would be
> happy to compare notes offline or over the phone.
>
> - Lisa
>
> [hidden email]
>
>
>
> -----Original Message-----
> From: Confocal Microscopy List on behalf of Bogdan Stoica
> Sent: Sun 8/23/2009 12:11 PM
> To: [hidden email]
> Subject: Re: [CONFOCALMICROSCOPY] Recommendations for commercial multi-photon
> system purchase
>  
> Hi fellows,
>
> I am also in the last stages of choosing a confocal/multiphoton system for
> our laboratory. After seeing multiple demos i am thorn between an Olympus
> FV1000MPE system and an Leica SP5 II. Our needs are to be able to do both
> confocal images on fixed and live in vitro cells and live in vivo rodent
> brain imaging. Both systems come with a UV, visible Argon four lines (488
> etc), laser diode 560 and 633 and IR laser (Chameleon for Leica and Maitai
> for Olympus), a complete set of Apo objectives as well as the famous 25x NA
> 1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes are
> inverted and the system also includes 2 NDD for Leica and 4 NDD in the case
> for Olympus.
> Fro what I could see the Leica software appeared more user friendly and I
> would give their system a better feel for confocal imaging and manufacturers
> potential upgrades- the system I am pricing does not include the AOBS
> feature, "white laser" or resonant scanner but those could be added later.
> On the other hand the Olympus seems a better choice for multiphoton imaging
> and an easier system to tinker with. The tech support and managers for both
> companies seemed outstanding to me here in the Washington DC-Baltimore area.
> I found interesting that I was not able to find any info on "how much" such
> a system should cost and after intense "negotiations" the quites range on my
> case around the ~700k mark with Olympus a little bellow and Leica a little
> above.
> I was wondering if people who recently had to make a similar decision can
> offer me some guidance both on the merits of their systems once they were
> installed as well as on the price, support etc.
> Thank you very much,
>
> Bogdan
>
>
>
> The information in this e-mail is intended only for the person to whom it is
> addressed. If you believe this e-mail was sent to you in error and the e-mail
> contains patient information, please contact the Partners Compliance HelpLine at
> http://www.partners.org/complianceline . If the e-mail was sent to you in error
> but does not contain patient information, please contact the sender and properly
> dispose of the e-mail.
>  

[Craig Brideau]

I've used the Leica white laser system during a demo.  It's nice, but
its main purpose is for core facilities where a user could walk in
with any arbitrary dye combination.  If you know what dyes you will be
using on a regular basis, just pick fixed laser lines to address them.
Regarding the multiphoton, if you are using a system that has an AOBS
or the like in it, you could have dispersion issues with the pulsed
laser.  I'd recommend getting one of the ones like the DeepSee with
built-in dispersion control in that case.
As for the objective lenses, apparently the 25x was designed
specifically for laser scanning applications, but I'd want to see a
side-by-side comparison with the 20x to be sure.

Craig

On Sun, Aug 23, 2009 at 2:03 PM, Bogdan Stoica<[hidden email]> wrote:

> Dear Lisa,
>
> Thank you for your kind response.
>  Let me adress the issue you raised in turn:
> -I did not get the chance to compare side by side the Olympus 25x vs. Leica
> 20x. I could only met with people that had rave reviews about the 25x so I
> would tend to five Olympus a slight advantage in this regard.
> -initially both systems had 4 NDDs but Leica had to leave only 2 to meet our
> budget. I saw people that having 4 NDDs coupled to 2 IR lasers and I was
> told me that even with one laser as in our case more NDDs could be
> beneficial for recording other channels such as intrinsic fluorescence etc.
> Again an extra point for Olympus.
> -You are right to point that for Leica features such as the AOBS as well as
> white laser should be unique advantages. The problem is that they would put
> the system outside our immediate budget although the fact that these
> upgrades are available in the future including as on-side upgrades for the
> AOBS justify an extra advantage for Leica.
> -Indeed, the need to do live cell imaging is directing us toward an inverted
> system.
> -the IR laser are the Maitai deepsee for olympus and the Chameleon Ultra II
> for the Leica.
>
> Bogdan
>

[Martin Hoppe-2]

COMMERCIAL VENDOR RESPONSE

Craig,

just to clarify: in the Leica TCS SP5, the Multiphoton laser does not pass the AOBS or AOTF. There is a separate dedicated IR beam splitter port. So there are no dispersion issues due to the pulsed IR beam passing through the acousto-optical tuneable elements.

Best regards
Martin

--------------------------------------------------------
Martin Hoppe, Ph.D.
Head of Market Management &
Confocal Application Management
Life Science Division

Leica Microsystems CMS GmbH
Am Friedensplatz 3 | 68165 Mannheim (Germany)
Phone : [phone] | Fax : [phone]
Cell: [phone]

I've used the Leica white laser system during a demo.  It's nice, but

its main purpose is for core facilities where a user could walk in

with any arbitrary dye combination.  If you know what dyes you will be

using on a regular basis, just pick fixed laser lines to address them.

Regarding the multiphoton, if you are using a system that has an AOBS

or the like in it, you could have dispersion issues with the pulsed

laser.  I'd recommend getting one of the ones like the DeepSee with

built-in dispersion control in that case.

As for the objective lenses, apparently the 25x was designed

specifically for laser scanning applications, but I'd want to see a

side-by-side comparison with the 20x to be sure.



Craig

-----Original Message-----
From: Craig Brideau <[hidden email]>
To: [hidden email]
Sent: Mon, Aug 24, 2009 5:02 pm
Subject: Re: Recommendations for commercial multi-photon system purchase

I've used the Leica white laser system during a demo.  It's nice, but

its main purpose is for core facilities where a user could walk in

with any arbitrary dye combination.  If you know what dyes you will be

using on a regular basis, just pick fixed laser lines to address them.

Regarding the multiphoton, if you are using a system that has an AOBS

or the like in it, you could have dispersion issues with the pulsed

laser.  I'd recommend getting one of the ones like the DeepSee with

built-in dispersion control in that case.

As for the objective lenses, apparently the 25x was designed

specifically for laser scanning applications, but I'd want to see a

side-by-side comparison with the 20x to be sure.



Craig



On Sun, Aug 23, 2009 at 2:03 PM, Bogdan Stoica<[hidden email]> wrote:

> Dear Lisa,

>

> Thank you for your kind response.

>  Let me adress the issue you raised in turn:

> -I did not get the chance to compare side by side the Olympus 25x vs. Leica

> 20x. I could only met with people that had rave reviews about the 25x so I

> would tend to five Olympus a slight advantage in this regard.

> -initially both systems had 4 NDDs but Leica had to leave only 2 to meet our

> budget. I saw people that having 4 NDDs coupled to 2 IR lasers and I was

> told me that even with one laser as in our case more NDDs could be

> beneficial for recording other channels such as intrinsic fluorescence etc.

> Again an extra point for Olympus.

> -You are right to point that for Leica features such as the AOBS as well as

> white laser should be unique advantages. The problem is that they would put

> the system outside our immediate budget although the fact that these

> upgrades are available in the future including as on-side upgrades for the

> AOBS justify an extra advantage for Leica.

> -Indeed, the need to do live cell imaging is directing us toward an inverted

> system.

> -the IR laser are the Maitai deepsee for olympus and the Chameleon Ultra II

> for the Leica.

>

> Bogdan

>

[Sylvie Le Guyader-2]

Hi Bogdan

We have recently gone through the purchase of a two photon/confocal
microscope. We opted for the Zeiss710 upright with a spectral detector and
for buying an extra confocal inverted for our cell work. When using an
inverted microscope, you offset the first plane you can image by the
thickness of the coverslip (160um) which is a real pity.

You must also consider that many lenses that are dedicated for 2 photon
imaging do not correct chromatic aberration in the visible range which means
that you will need another objective for confocal imaging with more than 1
fluorophore.

Does anyone know if Leica now supports femtosec pulsed lasers? They have so
far bypassed the Zeiss/Biorad patent by slowing down the laser to picosec
but I think the patent recent stopped. A psec laser would mean less
penetration which could be a problem for you on your inverted microscope,
especially if you want to image highly scattering samples.

Our experience (we have paid dearly before learning that) is that it is much
better to wait a bit longer for more money to come in and get a second
dedicated system. Piling up functions on one system often ends up
compromising all of them. Several companies (Zeiss, La Vision) offer
dedicated two photon systems that are a lot cheaper than two photon/confocal
combined and that are excellent for two photon imaging. If the system you
are looking at has both single and two photon lasers and if you also need
two sets of objectives, you are you are paying for all the expensive parts
twice anyway.

Good luck!


Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14157 Huddinge
Sweden
[phone]

>
> -----Original Message-----
> From: Confocal Microscopy List on behalf of Bogdan Stoica
> Sent: Sun 8/23/2009 12:11 PM
> To: [hidden email]
> Subject: Re: [CONFOCALMICROSCOPY] Recommendations for commercial multi-
> photon
> system purchase
>
> Hi fellows,
>
> I am also in the last stages of choosing a confocal/multiphoton system for
> our laboratory. After seeing multiple demos i am thorn between an Olympus
> FV1000MPE system and an Leica SP5 II. Our needs are to be able to do both
> confocal images on fixed and live in vitro cells and live in vivo rodent
> brain imaging. Both systems come with a UV, visible Argon four lines (488
> etc), laser diode 560 and 633 and IR laser (Chameleon for Leica and Maitai
> for Olympus), a complete set of Apo objectives as well as the famous 25x

NA
> 1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes are
> inverted and the system also includes 2 NDD for Leica and 4 NDD in the
case
> for Olympus.
> Fro what I could see the Leica software appeared more user friendly and I
> would give their system a better feel for confocal imaging and
manufacturers
> potential upgrades- the system I am pricing does not include the AOBS
> feature, "white laser" or resonant scanner but those could be added later.
> On the other hand the Olympus seems a better choice for multiphoton
imaging
> and an easier system to tinker with. The tech support and managers for
both
> companies seemed outstanding to me here in the Washington DC-Baltimore
area.
> I found interesting that I was not able to find any info on "how much"
such
> a system should cost and after intense "negotiations" the quites range on
my

> case around the ~700k mark with Olympus a little bellow and Leica a little
> above.
> I was wondering if people who recently had to make a similar decision can
> offer me some guidance both on the merits of their systems once they were
> installed as well as on the price, support etc.
> Thank you very much,
>
> Bogdan
>
>
>
> The information in this e-mail is intended only for the person to whom it

is
> addressed. If you believe this e-mail was sent to you in error and the
e-mail
> contains patient information, please contact the Partners Compliance
HelpLine at
> http://www.partners.org/complianceline . If the e-mail was sent to you in
error
> but does not contain patient information, please contact the sender and
properly
> dispose of the e-mail.

[Guy Cox]

It is entirely possible to avoid a coverslip on an inverted microscope - we just build a dam of dental moulding silicone around a dipping lens and fill it with water.  

We bought a Leica femtosecond 2-photon system back in 2000 - but the patent never applied to Australia. (That did mean it had to be put together here not in Germany).  My understanding is that Leica have now taken out a licence for what little time remains on the Cornell patent so that you can now buy a femtosecond system anywhere.  Actually you always could, you just had to do the integration yourself, since it was always the deal that no action would be taken against labs building a home-brew system.  But building a home-brew system isn't for everyone - and now there's no need for that.

I agree 100% that it is crazy to put every function under the sun on one scope - that's a recipe for huge conflicts of interest.

                                       Guy



Optical Imaging Techniques in Cell Biology
by Guy Cox    CRC Press / Taylor & Francis
    http://www.guycox.com/optical.htm
______________________________________________
Associate Professor Guy Cox, MA, DPhil(Oxon)
Electron Microscope Unit, Madsen Building F09,
University of Sydney, NSW 2006
______________________________________________
Phone [phone]     Fax [phone]
Mobile [phone]
______________________________________________
     http://www.guycox.net
-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Sylvie Le Guyader
Sent: Wednesday, 26 August 2009 7:10 PM
To: [hidden email]
Subject: Re: Recommendations for commercial multi-photon system purchase

Hi Bogdan

We have recently gone through the purchase of a two photon/confocal microscope. We opted for the Zeiss710 upright with a spectral detector and for buying an extra confocal inverted for our cell work. When using an inverted microscope, you offset the first plane you can image by the thickness of the coverslip (160um) which is a real pity.

You must also consider that many lenses that are dedicated for 2 photon imaging do not correct chromatic aberration in the visible range which means that you will need another objective for confocal imaging with more than 1 fluorophore.

Does anyone know if Leica now supports femtosec pulsed lasers? They have so far bypassed the Zeiss/Biorad patent by slowing down the laser to picosec but I think the patent recent stopped. A psec laser would mean less penetration which could be a problem for you on your inverted microscope, especially if you want to image highly scattering samples.

Our experience (we have paid dearly before learning that) is that it is much better to wait a bit longer for more money to come in and get a second dedicated system. Piling up functions on one system often ends up compromising all of them. Several companies (Zeiss, La Vision) offer dedicated two photon systems that are a lot cheaper than two photon/confocal combined and that are excellent for two photon imaging. If the system you are looking at has both single and two photon lasers and if you also need two sets of objectives, you are you are paying for all the expensive parts twice anyway.

Good luck!


Med vänlig hälsning / Best regards
 
Sylvie
 
@@@@@@@@@@@@@@@@@@@@@@@@
Sylvie Le Guyader
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14157 Huddinge
Sweden
[phone]

>
> -----Original Message-----
> From: Confocal Microscopy List on behalf of Bogdan Stoica
> Sent: Sun 8/23/2009 12:11 PM
> To: [hidden email]
> Subject: Re: [CONFOCALMICROSCOPY] Recommendations for commercial
> multi- photon system purchase
>
> Hi fellows,
>
> I am also in the last stages of choosing a confocal/multiphoton system
> for our laboratory. After seeing multiple demos i am thorn between an
> Olympus FV1000MPE system and an Leica SP5 II. Our needs are to be able
> to do both confocal images on fixed and live in vitro cells and live
> in vivo rodent brain imaging. Both systems come with a UV, visible
> Argon four lines (488 etc), laser diode 560 and 633 and IR laser
> (Chameleon for Leica and Maitai for Olympus), a complete set of Apo
> objectives as well as the famous 25x

NA
> 1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes
> are inverted and the system also includes 2 NDD for Leica and 4 NDD in
> the
case
> for Olympus.
> Fro what I could see the Leica software appeared more user friendly
> and I would give their system a better feel for confocal imaging and
manufacturers
> potential upgrades- the system I am pricing does not include the AOBS
> feature, "white laser" or resonant scanner but those could be added later.
> On the other hand the Olympus seems a better choice for multiphoton
imaging
> and an easier system to tinker with. The tech support and managers for
both
> companies seemed outstanding to me here in the Washington DC-Baltimore
area.
> I found interesting that I was not able to find any info on "how much"
such
> a system should cost and after intense "negotiations" the quites range
> on
my

> case around the ~700k mark with Olympus a little bellow and Leica a
> little above.
> I was wondering if people who recently had to make a similar decision
> can offer me some guidance both on the merits of their systems once
> they were installed as well as on the price, support etc.
> Thank you very much,
>
> Bogdan
>
>
>
> The information in this e-mail is intended only for the person to whom
> it

is
> addressed. If you believe this e-mail was sent to you in error and the
e-mail
> contains patient information, please contact the Partners Compliance
HelpLine at
> http://www.partners.org/complianceline . If the e-mail was sent to you
> in
error
> but does not contain patient information, please contact the sender
> and
properly
> dispose of the e-mail.

[Craig Brideau]

Thanks for the clarification!  It's good to know you have the
provision for a direct laser port.

Craig


On Wed, Aug 26, 2009 at 1:52 AM, <[hidden email]> wrote:

>
> COMMERCIAL VENDOR RESPONSE
>
> Craig,
>
> just to clarify: in the Leica TCS SP5, the Multiphoton laser does not pass
> the AOBS or AOTF. There is a separate dedicated IR beam splitter port. So
> there are no dispersion issues due to the pulsed IR beam passing through the
> acousto-optical tuneable elements.
>
> Best regards
> Martin
>
> --------------------------------------------------------
> Martin Hoppe, Ph.D.
> Head of Market Management &
> Confocal Application Management
> Life Science Division
>
> Leica Microsystems CMS GmbH
> Am Friedensplatz 3 | 68165 Mannheim (Germany)
> Phone : [phone] | Fax : [phone]
> Cell: [phone]
>
>
>
>
>
> I've used the Leica white laser system during a demo.  It's nice, but
>
> its main purpose is for core facilities where a user could walk in
>
> with any arbitrary dye combination.  If you know what dyes you will be
>
> using on a regular basis, just pick fixed laser lines to address them.
>
> Regarding the multiphoton, if you are using a system that has an AOBS
>
> or the like in it, you could have dispersion issues with the pulsed
>
> laser.  I'd recommend getting one of the ones like the DeepSee with
>
> built-in dispersion control in that case.
>
> As for the objective lenses, apparently the 25x was designed
>
> specifically for laser scanning applications, but I'd want to see a
>
> side-by-side comparison with the 20x to be sure.
>
>
>
> Craig
>
>
> -----Original Message-----
> From: Craig Brideau <[hidden email]>
> To: [hidden email]
> Sent: Mon, Aug 24, 2009 5:02 pm
> Subject: Re: Recommendations for commercial multi-photon system purchase
>
> I've used the Leica white laser system during a demo.  It's nice, but
>
> its main purpose is for core facilities where a user could walk in
>
> with any arbitrary dye combination.  If you know what dyes you will be
>
> using on a regular basis, just pick fixed laser lines to address them.
>
> Regarding the multiphoton, if you are using a system that has an AOBS
>
> or the like in it, you could have dispersion issues with the pulsed
>
> laser.  I'd recommend getting one of the ones like the DeepSee with
>
> built-in dispersion control in that case.
>
> As for the objective lenses, apparently the 25x was designed
>
> specifically for laser scanning applications, but I'd want to see a
>
> side-by-side comparison with the 20x to be sure.
>
>
>
> Craig
>
>
>
> On Sun, Aug 23, 2009 at 2:03 PM, Bogdan Stoica<[hidden email]>
> wrote:
>
>> Dear Lisa,
>
>>
>
>> Thank you for your kind response.
>
>>  Let me adress the issue you raised in turn:
>
>> -I did not get the chance to compare side by side the Olympus 25x vs.
>> Leica
>
>> 20x. I could only met with people that had rave reviews about the 25x so I
>
>> would tend to five Olympus a slight advantage in this regard.
>
>> -initially both systems had 4 NDDs but Leica had to leave only 2 to meet
>> our
>
>> budget. I saw people that having 4 NDDs coupled to 2 IR lasers and I was
>
>> told me that even with one laser as in our case more NDDs could be
>
>> beneficial for recording other channels such as intrinsic fluorescence
>> etc.
>
>> Again an extra point for Olympus.
>
>> -You are right to point that for Leica features such as the AOBS as well
>> as
>
>> white laser should be unique advantages. The problem is that they would
>> put
>
>> the system outside our immediate budget although the fact that these
>
>> upgrades are available in the future including as on-side upgrades for the
>
>> AOBS justify an extra advantage for Leica.
>
>> -Indeed, the need to do live cell imaging is directing us toward an
>> inverted
>
>> system.
>
>> -the IR laser are the Maitai deepsee for olympus and the Chameleon Ultra
>> II
>
>> for the Leica.
>
>>
>
>> Bogdan
>
>>
>
>

[Craig Brideau]

We have a semi-home-built combination 2-photon/confocal system which
saw a lot of use.  We bought a dedicated confocal (Nikon C1Si) to
offload the confocal users from the 2-photon system.  There was so
much demand for both modes that there were many scheduling conflicts.
It's much better to have dedicated systems if at all possible.

Craig

On Wed, Aug 26, 2009 at 6:27 AM, Guy Cox<[hidden email]> wrote:

> It is entirely possible to avoid a coverslip on an inverted microscope - we just build a dam of dental moulding silicone around a dipping lens and fill it with water.
>
> We bought a Leica femtosecond 2-photon system back in 2000 - but the patent never applied to Australia. (That did mean it had to be put together here not in Germany).  My understanding is that Leica have now taken out a licence for what little time remains on the Cornell patent so that you can now buy a femtosecond system anywhere.  Actually you always could, you just had to do the integration yourself, since it was always the deal that no action would be taken against labs building a home-brew system.  But building a home-brew system isn't for everyone - and now there's no need for that.
>
> I agree 100% that it is crazy to put every function under the sun on one scope - that's a recipe for huge conflicts of interest.
>
>                                       Guy
>
>
>
> Optical Imaging Techniques in Cell Biology
> by Guy Cox    CRC Press / Taylor & Francis
>    http://www.guycox.com/optical.htm
> ______________________________________________
> Associate Professor Guy Cox, MA, DPhil(Oxon)
> Electron Microscope Unit, Madsen Building F09,
> University of Sydney, NSW 2006
> ______________________________________________
> Phone [phone]     Fax [phone]
> Mobile [phone]
> ______________________________________________
>     http://www.guycox.net
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Sylvie Le Guyader
> Sent: Wednesday, 26 August 2009 7:10 PM
> To: [hidden email]
> Subject: Re: Recommendations for commercial multi-photon system purchase
>
> Hi Bogdan
>
> We have recently gone through the purchase of a two photon/confocal microscope. We opted for the Zeiss710 upright with a spectral detector and for buying an extra confocal inverted for our cell work. When using an inverted microscope, you offset the first plane you can image by the thickness of the coverslip (160um) which is a real pity.
>
> You must also consider that many lenses that are dedicated for 2 photon imaging do not correct chromatic aberration in the visible range which means that you will need another objective for confocal imaging with more than 1 fluorophore.
>
> Does anyone know if Leica now supports femtosec pulsed lasers? They have so far bypassed the Zeiss/Biorad patent by slowing down the laser to picosec but I think the patent recent stopped. A psec laser would mean less penetration which could be a problem for you on your inverted microscope, especially if you want to image highly scattering samples.
>
> Our experience (we have paid dearly before learning that) is that it is much better to wait a bit longer for more money to come in and get a second dedicated system. Piling up functions on one system often ends up compromising all of them. Several companies (Zeiss, La Vision) offer dedicated two photon systems that are a lot cheaper than two photon/confocal combined and that are excellent for two photon imaging. If the system you are looking at has both single and two photon lasers and if you also need two sets of objectives, you are you are paying for all the expensive parts twice anyway.
>
> Good luck!
>
>
> Med vänlig hälsning / Best regards
>
> Sylvie
>
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader
> Dept of Biosciences and Nutrition
> Karolinska Institutet
> Novum
> 14157 Huddinge
> Sweden
> [phone]
>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List on behalf of Bogdan Stoica
>> Sent: Sun 8/23/2009 12:11 PM
>> To: [hidden email]
>> Subject: Re: [CONFOCALMICROSCOPY] Recommendations for commercial
>> multi- photon system purchase
>>
>> Hi fellows,
>>
>> I am also in the last stages of choosing a confocal/multiphoton system
>> for our laboratory. After seeing multiple demos i am thorn between an
>> Olympus FV1000MPE system and an Leica SP5 II. Our needs are to be able
>> to do both confocal images on fixed and live in vitro cells and live
>> in vivo rodent brain imaging. Both systems come with a UV, visible
>> Argon four lines (488 etc), laser diode 560 and 633 and IR laser
>> (Chameleon for Leica and Maitai for Olympus), a complete set of Apo
>> objectives as well as the famous 25x
> NA
>> 1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes
>> are inverted and the system also includes 2 NDD for Leica and 4 NDD in
>> the
> case
>> for Olympus.
>> Fro what I could see the Leica software appeared more user friendly
>> and I would give their system a better feel for confocal imaging and
> manufacturers
>> potential upgrades- the system I am pricing does not include the AOBS
>> feature, "white laser" or resonant scanner but those could be added later.
>> On the other hand the Olympus seems a better choice for multiphoton
> imaging
>> and an easier system to tinker with. The tech support and managers for
> both
>> companies seemed outstanding to me here in the Washington DC-Baltimore
> area.
>> I found interesting that I was not able to find any info on "how much"
> such
>> a system should cost and after intense "negotiations" the quites range
>> on
> my
>> case around the ~700k mark with Olympus a little bellow and Leica a
>> little above.
>> I was wondering if people who recently had to make a similar decision
>> can offer me some guidance both on the merits of their systems once
>> they were installed as well as on the price, support etc.
>> Thank you very much,
>>
>> Bogdan
>>
>>
>>
>> The information in this e-mail is intended only for the person to whom
>> it
> is
>> addressed. If you believe this e-mail was sent to you in error and the
> e-mail
>> contains patient information, please contact the Partners Compliance
> HelpLine at
>> http://www.partners.org/complianceline . If the e-mail was sent to you
>> in
> error
>> but does not contain patient information, please contact the sender
>> and
> properly
>> dispose of the e-mail.
>

[neeraj Gohad-3]

Hi Craig,

 

We are thinking of building a semi-home built 2-photon confocal system ourselves. Could you please tell me a little more about your system.

 

Thanks,

 

Neeraj.

 

 

 

 

Neeraj V. Gohad, Ph.D.

Postdoctoral Fellow

Okeanos Research Group

Department of Biological Sciences

132 Long Hall

Clemson University

Clemson,SC-29634

Phone: [phone]

Fax: [phone]

 

 

 

 

 

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Craig Brideau
Sent: Wednesday, August 26, 2009 10:58 AM
To: [hidden email]
Subject: Re: Recommendations for commercial multi-photon system purchase

 

We have a semi-home-built combination 2-photon/confocal system which

saw a lot of use.  We bought a dedicated confocal (Nikon C1Si) to

offload the confocal users from the 2-photon system.  There was so

much demand for both modes that there were many scheduling conflicts.

It's much better to have dedicated systems if at all possible.

 

Craig

 

On Wed, Aug 26, 2009 at 6:27 AM, Guy Cox<[hidden email]> wrote:

> It is entirely possible to avoid a coverslip on an inverted microscope - we just build a dam of dental moulding silicone around a dipping lens and fill it with water.

> 

> We bought a Leica femtosecond 2-photon system back in 2000 - but the patent never applied to Australia. (That did mean it had to be put together here not in Germany).  My understanding is that Leica have now taken out a licence for what little time remains on the Cornell patent so that you can now buy a femtosecond system anywhere.  Actually you always could, you just had to do the integration yourself, since it was always the deal that no action would be taken against labs building a home-brew system.  But building a home-brew system isn't for everyone - and now there's no need for that.

> 

> I agree 100% that it is crazy to put every function under the sun on one scope - that's a recipe for huge conflicts of interest.

> 

>                                       Guy

> 

> 

> 

> Optical Imaging Techniques in Cell Biology

> by Guy Cox    CRC Press / Taylor & Francis

>    http://www.guycox.com/optical.htm

> ______________________________________________

> Associate Professor Guy Cox, MA, DPhil(Oxon)

> Electron Microscope Unit, Madsen Building F09,

> University of Sydney, NSW 2006

> ______________________________________________

> Phone [phone]     Fax [phone]

> Mobile [phone]

> ______________________________________________

>     http://www.guycox.net

> -----Original Message-----

> From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Sylvie Le Guyader

> Sent: Wednesday, 26 August 2009 7:10 PM

> To: [hidden email]

> Subject: Re: Recommendations for commercial multi-photon system purchase

> 

> Hi Bogdan

> 

> We have recently gone through the purchase of a two photon/confocal microscope. We opted for the Zeiss710 upright with a spectral detector and for buying an extra confocal inverted for our cell work. When using an inverted microscope, you offset the first plane you can image by the thickness of the coverslip (160um) which is a real pity.

> 

> You must also consider that many lenses that are dedicated for 2 photon imaging do not correct chromatic aberration in the visible range which means that you will need another objective for confocal imaging with more than 1 fluorophore.

> 

> Does anyone know if Leica now supports femtosec pulsed lasers? They have so far bypassed the Zeiss/Biorad patent by slowing down the laser to picosec but I think the patent recent stopped. A psec laser would mean less penetration which could be a problem for you on your inverted microscope, especially if you want to image highly scattering samples.

> 

> Our experience (we have paid dearly before learning that) is that it is much better to wait a bit longer for more money to come in and get a second dedicated system. Piling up functions on one system often ends up compromising all of them. Several companies (Zeiss, La Vision) offer dedicated two photon systems that are a lot cheaper than two photon/confocal combined and that are excellent for two photon imaging. If the system you are looking at has both single and two photon lasers and if you also need two sets of objectives, you are you are paying for all the expensive parts twice anyway.

> 

> Good luck!

> 

> 

> Med vänlig hälsning / Best regards

> 

> Sylvie

> 

> @@@@@@@@@@@@@@@@@@@@@@@@

> Sylvie Le Guyader

> Dept of Biosciences and Nutrition

> Karolinska Institutet

> Novum

> 14157 Huddinge

> Sweden

> [phone]

> 

>> 

>> -----Original Message-----

>> From: Confocal Microscopy List on behalf of Bogdan Stoica

>> Sent: Sun 8/23/2009 12:11 PM

>> To: [hidden email]

>> Subject: Re: [CONFOCALMICROSCOPY] Recommendations for commercial

>> multi- photon system purchase

>> 

>> Hi fellows,

>> 

>> I am also in the last stages of choosing a confocal/multiphoton system

>> for our laboratory. After seeing multiple demos i am thorn between an

>> Olympus FV1000MPE system and an Leica SP5 II. Our needs are to be able

>> to do both confocal images on fixed and live in vitro cells and live

>> in vivo rodent brain imaging. Both systems come with a UV, visible

>> Argon four lines (488 etc), laser diode 560 and 633 and IR laser

>> (Chameleon for Leica and Maitai for Olympus), a complete set of Apo

>> objectives as well as the famous 25x

> NA

>> 1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes

>> are inverted and the system also includes 2 NDD for Leica and 4 NDD in

>> the

> case

>> for Olympus.

>> Fro what I could see the Leica software appeared more user friendly

>> and I would give their system a better feel for confocal imaging and

> manufacturers

>> potential upgrades- the system I am pricing does not include the AOBS

>> feature, "white laser" or resonant scanner but those could be added later.

>> On the other hand the Olympus seems a better choice for multiphoton

> imaging

>> and an easier system to tinker with. The tech support and managers for

> both

>> companies seemed outstanding to me here in the Washington DC-Baltimore

> area.

>> I found interesting that I was not able to find any info on "how much"

> such

>> a system should cost and after intense "negotiations" the quites range

>> on

> my

>> case around the ~700k mark with Olympus a little bellow and Leica a

>> little above.

>> I was wondering if people who recently had to make a similar decision

>> can offer me some guidance both on the merits of their systems once

>> they were installed as well as on the price, support etc.

>> Thank you very much,

>> 

>> Bogdan

>> 

>> 

>> 

>> The information in this e-mail is intended only for the person to whom

>> it

> is

>> addressed. If you believe this e-mail was sent to you in error and the

> e-mail

>> contains patient information, please contact the Partners Compliance

> HelpLine at

>> http://www.partners.org/complianceline . If the e-mail was sent to you

>> in

> error

>> but does not contain patient information, please contact the sender

>> and

> properly

>> dispose of the e-mail.

> 

[Armstrong, Brian]

Dr Hoppe, does Leica offer an objective that rivals the Zeiss 20x/1.0 and the Olympus 25x multiphoton objectives?

Can you share the specifics? (NA, thread size, w.d., parfocal length, etc..)

Thanks,

 

Brian D Armstrong PhD

Light Microscopy Core Manager

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

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of [hidden email]
Sent: Wednesday, August 26, 2009 12:53 AM
To: [hidden email]
Subject: Re: Recommendations for commercial multi-photon system purchase

 

COMMERCIAL VENDOR RESPONSE

Craig,

just to clarify: in the Leica TCS SP5, the Multiphoton laser does not pass the AOBS or AOTF. There is a separate dedicated IR beam splitter port. So there are no dispersion issues due to the pulsed IR beam passing through the acousto-optical tuneable elements.

Best regards
Martin

--------------------------------------------------------
Martin Hoppe, Ph.D.
Head of Market Management &
Confocal Application Management
Life Science Division

Leica Microsystems CMS GmbH
Am Friedensplatz 3 | 68165 Mannheim (Germany)
Phone : [phone] | Fax : [phone]
Cell: [phone]

I've used the Leica white laser system during a demo.  It's nice, but

its main purpose is for core facilities where a user could walk in

with any arbitrary dye combination.  If you know what dyes you will be

using on a regular basis, just pick fixed laser lines to address them.

Regarding the multiphoton, if you are using a system that has an AOBS

or the like in it, you could have dispersion issues with the pulsed

laser.  I'd recommend getting one of the ones like the DeepSee with

built-in dispersion control in that case.

As for the objective lenses, apparently the 25x was designed

specifically for laser scanning applications, but I'd want to see a

side-by-side comparison with the 20x to be sure.

Craig

 

 

 

-----Original Message-----
From: Craig Brideau <[hidden email]>
To: [hidden email]
Sent: Mon, Aug 24, 2009 5:02 pm
Subject: Re: Recommendations for commercial multi-photon system purchase

I've used the Leica white laser system during a demo.  It's nice, but

its main purpose is for core facilities where a user could walk in

with any arbitrary dye combination.  If you know what dyes you will be

using on a regular basis, just pick fixed laser lines to address them.

Regarding the multiphoton, if you are using a system that has an AOBS

or the like in it, you could have dispersion issues with the pulsed

laser.  I'd recommend getting one of the ones like the DeepSee with

built-in dispersion control in that case.

As for the objective lenses, apparently the 25x was designed

specifically for laser scanning applications, but I'd want to see a

side-by-side comparison with the 20x to be sure.

Craig

On Sun, Aug 23, 2009 at 2:03 PM, Bogdan Stoica<[hidden email]> wrote:

> Dear Lisa,

>

> Thank you for your kind response.

>  Let me adress the issue you raised in turn:

> -I did not get the chance to compare side by side the Olympus 25x vs. Leica

> 20x. I could only met with people that had rave reviews about the 25x so I

> would tend to five Olympus a slight advantage in this regard.

> -initially both systems had 4 NDDs but Leica had to leave only 2 to meet our

> budget. I saw people that having 4 NDDs coupled to 2 IR lasers and I was

> told me that even with one laser as in our case more NDDs could be

> beneficial for recording other channels such as intrinsic fluorescence etc.

> Again an extra point for Olympus.

> -You are right to point that for Leica features such as the AOBS as well as

> white laser should be unique advantages. The problem is that they would put

> the system outside our immediate budget although the fact that these

> upgrades are available in the future including as on-side upgrades for the

> AOBS justify an extra advantage for Leica.

> -Indeed, the need to do live cell imaging is directing us toward an inverted

> system.

> -the IR laser are the Maitai deepsee for olympus and the Chameleon Ultra II

> for the Leica.

>

> Bogdan

>

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[Craig Brideau]

We use a Spectra Physics Tsunami.  It's all manual so it can be tricky
to use, but the lack of motorization and simplicity of the design
seems to have given it a long lifespan.  The lab bought it in ~2001,
and I inherited the care and maintenance of the system when I joined
the lab a few years ago.  The laser is seven or eight years old now
and still quite happily chugging along.  I've even managed to teach
the biology people how to tune it themselves @;-).  The scan head had
the galvos wear out once (it saw a LOT of use) and we had to replace
them, but since then it's still a regular performer for our lab.  You
can see the paper my predecessor published on the build:

Conversion of the Nikon C1 Confocal Laser-Scanning Head for
Multiphoton Excitation on an Upright Microscope
Andrew Ridsdale, Ileana Micu, and Peter K. Stys
Applied Optics, Vol. 43, Issue 8, pp. 1669-1675        doi:10.1364/AO.43.001669

Find it at:

http://www.opticsinfobase.org/abstract.cfm?URI=ao-43-8-1669

Hope you guys find it useful!

Craig



On Wed, Aug 26, 2009 at 9:58 AM, Sergey Egorov<[hidden email]> wrote:

> Hi Craig,
>
> so what femtosecond laser you use in your semi-home-built system?
>
> here is one home-brew multiphoton system built at UCLA based on our Trestles
> lasers http://www.dmphotonics.com/trestles%20brochure%20web.pdf
>
> http://www.youtube.com/watch?v=b0jmCqoHqkc&feature=channel_page
>
> Best regards,
> Sergey Egorov
> Del Mar Photonics
> 4119 Twilight Ridge
> San Diego, CA 92130
> tel [phone]
> fax [phone]
> www.dmphotonics.com
> [hidden email]
>
>> We have a semi-home-built combination 2-photon/confocal system which
>> saw a lot of use.  We bought a dedicated confocal (Nikon C1Si) to
>> offload the confocal users from the 2-photon system.  There was so
>> much demand for both modes that there were many scheduling conflicts.
>> It's much better to have dedicated systems if at all possible.
>>
>> Craig
>>
>> On Wed, Aug 26, 2009 at 6:27 AM, Guy Cox<[hidden email]> wrote:
>>
>>>
>>> It is entirely possible to avoid a coverslip on an inverted microscope -
>>> we just build a dam of dental moulding silicone around a dipping lens and
>>> fill it with water.
>>>
>>> We bought a Leica femtosecond 2-photon system back in 2000 - but the
>>> patent never applied to Australia. (That did mean it had to be put together
>>> here not in Germany).  My understanding is that Leica have now taken out a
>>> licence for what little time remains on the Cornell patent so that you can
>>> now buy a femtosecond system anywhere.  Actually you always could, you just
>>> had to do the integration yourself, since it was always the deal that no
>>> action would be taken against labs building a home-brew system.  But
>>> building a home-brew system isn't for everyone - and now there's no need for
>>> that.
>>>
>>> I agree 100% that it is crazy to put every function under the sun on one
>>> scope - that's a recipe for huge conflicts of interest.
>>>
>>>                                      Guy
>>>
>>>
>>>
>>> Optical Imaging Techniques in Cell Biology
>>> by Guy Cox    CRC Press / Taylor & Francis
>>>   http://www.guycox.com/optical.htm
>>> ______________________________________________
>>> Associate Professor Guy Cox, MA, DPhil(Oxon)
>>> Electron Microscope Unit, Madsen Building F09,
>>> University of Sydney, NSW 2006
>>> ______________________________________________
>>> Phone [phone]     Fax [phone]
>>> Mobile [phone]
>>> ______________________________________________
>>>    http://www.guycox.net
>>> -----Original Message-----
>>> From: Confocal Microscopy List [mailto:[hidden email]]
>>> On Behalf Of Sylvie Le Guyader
>>> Sent: Wednesday, 26 August 2009 7:10 PM
>>> To: [hidden email]
>>> Subject: Re: Recommendations for commercial multi-photon system purchase
>>>
>>> Hi Bogdan
>>>
>>> We have recently gone through the purchase of a two photon/confocal
>>> microscope. We opted for the Zeiss710 upright with a spectral detector and
>>> for buying an extra confocal inverted for our cell work. When using an
>>> inverted microscope, you offset the first plane you can image by the
>>> thickness of the coverslip (160um) which is a real pity.
>>>
>>> You must also consider that many lenses that are dedicated for 2 photon
>>> imaging do not correct chromatic aberration in the visible range which means
>>> that you will need another objective for confocal imaging with more than 1
>>> fluorophore.
>>>
>>> Does anyone know if Leica now supports femtosec pulsed lasers? They have
>>> so far bypassed the Zeiss/Biorad patent by slowing down the laser to picosec
>>> but I think the patent recent stopped. A psec laser would mean less
>>> penetration which could be a problem for you on your inverted microscope,
>>> especially if you want to image highly scattering samples.
>>>
>>> Our experience (we have paid dearly before learning that) is that it is
>>> much better to wait a bit longer for more money to come in and get a second
>>> dedicated system. Piling up functions on one system often ends up
>>> compromising all of them. Several companies (Zeiss, La Vision) offer
>>> dedicated two photon systems that are a lot cheaper than two photon/confocal
>>> combined and that are excellent for two photon imaging. If the system you
>>> are looking at has both single and two photon lasers and if you also need
>>> two sets of objectives, you are you are paying for all the expensive parts
>>> twice anyway.
>>>
>>> Good luck!
>>>
>>>
>>> Med vänlig hälsning / Best regards
>>>
>>> Sylvie
>>>
>>> @@@@@@@@@@@@@@@@@@@@@@@@
>>> Sylvie Le Guyader
>>> Dept of Biosciences and Nutrition
>>> Karolinska Institutet
>>> Novum
>>> 14157 Huddinge
>>> Sweden
>>> [phone]
>>>
>>>
>>>>
>>>> -----Original Message-----
>>>> From: Confocal Microscopy List on behalf of Bogdan Stoica
>>>> Sent: Sun 8/23/2009 12:11 PM
>>>> To: [hidden email]
>>>> Subject: Re: [CONFOCALMICROSCOPY] Recommendations for commercial
>>>> multi- photon system purchase
>>>>
>>>> Hi fellows,
>>>>
>>>> I am also in the last stages of choosing a confocal/multiphoton system
>>>> for our laboratory. After seeing multiple demos i am thorn between an
>>>> Olympus FV1000MPE system and an Leica SP5 II. Our needs are to be able
>>>> to do both confocal images on fixed and live in vitro cells and live
>>>> in vivo rodent brain imaging. Both systems come with a UV, visible
>>>> Argon four lines (488 etc), laser diode 560 and 633 and IR laser
>>>> (Chameleon for Leica and Maitai for Olympus), a complete set of Apo
>>>> objectives as well as the famous 25x
>>>>
>>>
>>> NA
>>>
>>>>
>>>> 1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes
>>>> are inverted and the system also includes 2 NDD for Leica and 4 NDD in
>>>> the
>>>>
>>>
>>> case
>>>
>>>>
>>>> for Olympus.
>>>> Fro what I could see the Leica software appeared more user friendly
>>>> and I would give their system a better feel for confocal imaging and
>>>>
>>>
>>> manufacturers
>>>
>>>>
>>>> potential upgrades- the system I am pricing does not include the AOBS
>>>> feature, "white laser" or resonant scanner but those could be added
>>>> later.
>>>> On the other hand the Olympus seems a better choice for multiphoton
>>>>
>>>
>>> imaging
>>>
>>>>
>>>> and an easier system to tinker with. The tech support and managers for
>>>>
>>>
>>> both
>>>
>>>>
>>>> companies seemed outstanding to me here in the Washington DC-Baltimore
>>>>
>>>
>>> area.
>>>
>>>>
>>>> I found interesting that I was not able to find any info on "how much"
>>>>
>>>
>>> such
>>>
>>>>
>>>> a system should cost and after intense "negotiations" the quites range
>>>> on
>>>>
>>>
>>> my
>>>
>>>>
>>>> case around the ~700k mark with Olympus a little bellow and Leica a
>>>> little above.
>>>> I was wondering if people who recently had to make a similar decision
>>>> can offer me some guidance both on the merits of their systems once
>>>> they were installed as well as on the price, support etc.
>>>> Thank you very much,
>>>>
>>>> Bogdan
>>>>
>>>>
>>>>
>>>> The information in this e-mail is intended only for the person to whom
>>>> it
>>>>
>>>
>>> is
>>>
>>>>
>>>> addressed. If you believe this e-mail was sent to you in error and the
>>>>
>>>
>>> e-mail
>>>
>>>>
>>>> contains patient information, please contact the Partners Compliance
>>>>
>>>
>>> HelpLine at
>>>
>>>>
>>>> http://www.partners.org/complianceline . If the e-mail was sent to you
>>>> in
>>>>
>>>
>>> error
>>>
>>>>
>>>> but does not contain patient information, please contact the sender
>>>> and
>>>>
>>>
>>> properly
>>>
>>>>
>>>> dispose of the e-mail.
>>>>
>>
>>
>
>
>

[Rosemary.White]

Hear hear!  Sylvie's recommendation to purchase multiple (e.g. two)
dedicated systems rather than try to make one system do everything for
everyone is spot-on.  Getting a single multi-function system is always a
compromise, especially these days when there are specialist objectives,
light paths etc. for specific applications.  And especially if you have a
central facility - either support them with funding or try to get the
funding for dedicated, specialist confocals.  Apart from the compromises,
you also find that a single confocal is booked up to its eyeballs within
minutes of installation...

Rosemary

Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

ph 61 2 6246 5475
fx 61 2 6246 5334



On 26/08/09 7:09 PM, "Sylvie Le Guyader" <[hidden email]> wrote:

> Hi Bogdan
>
> We have recently gone through the purchase of a two photon/confocal
> microscope. We opted for the Zeiss710 upright with a spectral detector and
> for buying an extra confocal inverted for our cell work. When using an
> inverted microscope, you offset the first plane you can image by the
> thickness of the coverslip (160um) which is a real pity.
>
> You must also consider that many lenses that are dedicated for 2 photon
> imaging do not correct chromatic aberration in the visible range which means
> that you will need another objective for confocal imaging with more than 1
> fluorophore.
>
> Does anyone know if Leica now supports femtosec pulsed lasers? They have so
> far bypassed the Zeiss/Biorad patent by slowing down the laser to picosec
> but I think the patent recent stopped. A psec laser would mean less
> penetration which could be a problem for you on your inverted microscope,
> especially if you want to image highly scattering samples.
>
> Our experience (we have paid dearly before learning that) is that it is much
> better to wait a bit longer for more money to come in and get a second
> dedicated system. Piling up functions on one system often ends up
> compromising all of them. Several companies (Zeiss, La Vision) offer
> dedicated two photon systems that are a lot cheaper than two photon/confocal
> combined and that are excellent for two photon imaging. If the system you
> are looking at has both single and two photon lasers and if you also need
> two sets of objectives, you are you are paying for all the expensive parts
> twice anyway.
>
> Good luck!
>
>
> Med vänlig hälsning / Best regards
>  
> Sylvie
>  
> @@@@@@@@@@@@@@@@@@@@@@@@
> Sylvie Le Guyader
> Dept of Biosciences and Nutrition
> Karolinska Institutet
> Novum
> 14157 Huddinge
> Sweden
> [phone]
>
>>
>> -----Original Message-----
>> From: Confocal Microscopy List on behalf of Bogdan Stoica
>> Sent: Sun 8/23/2009 12:11 PM
>> To: [hidden email]
>> Subject: Re: [CONFOCALMICROSCOPY] Recommendations for commercial multi-
>> photon
>> system purchase
>>
>> Hi fellows,
>>
>> I am also in the last stages of choosing a confocal/multiphoton system for
>> our laboratory. After seeing multiple demos i am thorn between an Olympus
>> FV1000MPE system and an Leica SP5 II. Our needs are to be able to do both
>> confocal images on fixed and live in vitro cells and live in vivo rodent
>> brain imaging. Both systems come with a UV, visible Argon four lines (488
>> etc), laser diode 560 and 633 and IR laser (Chameleon for Leica and Maitai
>> for Olympus), a complete set of Apo objectives as well as the famous 25x
> NA
>> 1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes are
>> inverted and the system also includes 2 NDD for Leica and 4 NDD in the
> case
>> for Olympus.
>> Fro what I could see the Leica software appeared more user friendly and I
>> would give their system a better feel for confocal imaging and
> manufacturers
>> potential upgrades- the system I am pricing does not include the AOBS
>> feature, "white laser" or resonant scanner but those could be added later.
>> On the other hand the Olympus seems a better choice for multiphoton
> imaging
>> and an easier system to tinker with. The tech support and managers for
> both
>> companies seemed outstanding to me here in the Washington DC-Baltimore
> area.
>> I found interesting that I was not able to find any info on "how much"
> such
>> a system should cost and after intense "negotiations" the quites range on
> my
>> case around the ~700k mark with Olympus a little bellow and Leica a little
>> above.
>> I was wondering if people who recently had to make a similar decision can
>> offer me some guidance both on the merits of their systems once they were
>> installed as well as on the price, support etc.
>> Thank you very much,
>>
>> Bogdan
>>
>>
>>
>> The information in this e-mail is intended only for the person to whom it
> is
>> addressed. If you believe this e-mail was sent to you in error and the
> e-mail
>> contains patient information, please contact the Partners Compliance
> HelpLine at
>> http://www.partners.org/complianceline . If the e-mail was sent to you in
> error
>> but does not contain patient information, please contact the sender and
> properly
>> dispose of the e-mail.

[Martin Hoppe-2]

Brian,

 

There is a Leica PLAN APO HCX 20x 1,0 NA Water immersion Objective with 2 mm Free Working Distance, optimized for Multiphoton Microscopy and Electrophysiology.

 

Your local Leica Representative can share further details with you.

 

Best regards

Martin

 

In a message dated 26.08.2009 19:14:59 Mitteleuropäische Sommerzeit, [hidden email] writes:

Dr Hoppe, does Leica offer an objective that rivals the Zeiss 20x/1.0 and the Olympus 25x multiphoton objectives?

Can you share the specifics? (NA, thread size, w.d., parfocal length, etc..)

Thanks,

 

Brian D Armstrong PhD

Light Microscopy Core Manager

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

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of [hidden email]
Sent: Wednesday, August 26, 2009 12:53 AM
To: [hidden email]
Subject: Re: Recommendations for commercial multi-photon system purchase

 

COMMERCIAL VENDOR RESPONSE

Craig,

just to clarify: in the Leica TCS SP5, the Multiphoton laser does not pass the AOBS or AOTF. There is a separate dedicated IR beam splitter port. So there are no dispersion issues due to the pulsed IR beam passing through the acousto-optical tuneable elements.

Best regards
Martin

--------------------------------------------------------
Martin Hoppe, Ph.D.
Head of Market Management &
Confocal Application Management
Life Science Division

Leica Microsystems CMS GmbH
Am Friedensplatz 3 | 68165 Mannheim (Germany)
Phone : [phone] | Fax : [phone]
Cell: [phone]

I've used the Leica white laser system during a demo.  It's nice, but

its main purpose is for core facilities where a user could walk in

with any arbitrary dye combination.  If you know what dyes you will be

using on a regular basis, just pick fixed laser lines to address them.

Regarding the multiphoton, if you are using a system that has an AOBS

or the like in it, you could have dispersion issues with the pulsed

laser.  I'd recommend getting one of the ones like the DeepSee with

built-in dispersion control in that case.

As for the objective lenses, apparently the 25x was designed

specifically for laser scanning applications, but I'd want to see a

side-by-side comparison with the 20x to be sure.

Craig

 

 

 

-----Original Message-----
From: Craig Brideau <[hidden email]>
To: [hidden email]
Sent: Mon, Aug 24, 2009 5:02 pm
Subject: Re: Recommendations for commercial multi-photon system purchase

I've used the Leica white laser system during a demo.  It's nice, but

its main purpose is for core facilities where a user could walk in

with any arbitrary dye combination.  If you know what dyes you will be

using on a regular basis, just pick fixed laser lines to address them.

Regarding the multiphoton, if you are using a system that has an AOBS

or the like in it, you could have dispersion issues with the pulsed

laser.  I'd recommend getting one of the ones like the DeepSee with

built-in dispersion control in that case.

As for the objective lenses, apparently the 25x was designed

specifically for laser scanning applications, but I'd want to see a

side-by-side comparison with the 20x to be sure.

Craig

On Sun, Aug 23, 2009 at 2:03 PM, Bogdan Stoica<[hidden email]> wrote:

> Dear Lisa,

>

> Thank you for your kind response.

>  Let me adress the issue you raised in turn:

> -I did not get the chance to compare side by side the Olympus 25x vs. Leica

> 20x. I could only met with people that had rave reviews about the 25x so I

> would tend to five Olympus a slight advantage in this regard.

> -initially both systems had 4 NDDs but Leica had to leave only 2 to meet our

> budget. I saw people that having 4 NDDs coupled to 2 IR lasers and I was

> told me that even with one laser as in our case more NDDs could be

> beneficial for recording other channels such as intrinsic fluorescence etc.

> Again an extra point for Olympus.

> -You are right to point that for Leica features such as the AOBS as well as

> white laser should be unique advantages. The problem is that they would put

> the system outside our immediate budget although the fact that these

> upgrades are available in the future including as on-side upgrades for the

> AOBS justify an extra advantage for Leica.

> -Indeed, the need to do live cell imaging is directing us toward an inverted

> system.

> -the IR laser are the Maitai deepsee for olympus and the Chameleon Ultra II

> for the Leica.

>

> Bogdan

>

---------------------------------------------------------------------
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[Armstrong, Brian]

Thank you.

 

Brian D Armstrong PhD

Light Microscopy Core Manager

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

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of [hidden email]
Sent: Thursday, August 27, 2009 11:50 AM
To: [hidden email]
Subject: Re: Recommendations for commercial multi-photon system purchase

 

Brian,

 

There is a Leica PLAN APO HCX 20x 1,0 NA Water immersion Objective with 2 mm Free Working Distance, optimized for Multiphoton Microscopy and Electrophysiology.

 

Your local Leica Representative can share further details with you.

 

Best regards

Martin

 

In a message dated 26.08.2009 19:14:59 Mitteleuropäische Sommerzeit, [hidden email] writes:

Dr Hoppe, does Leica offer an objective that rivals the Zeiss 20x/1.0 and the Olympus 25x multiphoton objectives?

Can you share the specifics? (NA, thread size, w.d., parfocal length, etc..)

Thanks,

 

Brian D Armstrong PhD

Light Microscopy Core Manager

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

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of [hidden email]
Sent: Wednesday, August 26, 2009 12:53 AM
To: [hidden email]
Subject: Re: Recommendations for commercial multi-photon system purchase

 

COMMERCIAL VENDOR RESPONSE

Craig,

just to clarify: in the Leica TCS SP5, the Multiphoton laser does not pass the AOBS or AOTF. There is a separate dedicated IR beam splitter port. So there are no dispersion issues due to the pulsed IR beam passing through the acousto-optical tuneable elements.

Best regards
Martin

--------------------------------------------------------
Martin Hoppe, Ph.D.
Head of Market Management &
Confocal Application Management
Life Science Division

Leica Microsystems CMS GmbH
Am Friedensplatz 3 | 68165 Mannheim (Germany)
Phone : [phone] | Fax : [phone]
Cell: [phone]

I've used the Leica white laser system during a demo.  It's nice, but

its main purpose is for core facilities where a user could walk in

with any arbitrary dye combination.  If you know what dyes you will be

using on a regular basis, just pick fixed laser lines to address them.

Regarding the multiphoton, if you are using a system that has an AOBS

or the like in it, you could have dispersion issues with the pulsed

laser.  I'd recommend getting one of the ones like the DeepSee with

built-in dispersion control in that case.

As for the objective lenses, apparently the 25x was designed

specifically for laser scanning applications, but I'd want to see a

side-by-side comparison with the 20x to be sure.

Craig

 

 

 

-----Original Message-----
From: Craig Brideau <[hidden email]>
To: [hidden email]
Sent: Mon, Aug 24, 2009 5:02 pm
Subject: Re: Recommendations for commercial multi-photon system purchase

I've used the Leica white laser system during a demo.  It's nice, but

its main purpose is for core facilities where a user could walk in

with any arbitrary dye combination.  If you know what dyes you will be

using on a regular basis, just pick fixed laser lines to address them.

Regarding the multiphoton, if you are using a system that has an AOBS

or the like in it, you could have dispersion issues with the pulsed

laser.  I'd recommend getting one of the ones like the DeepSee with

built-in dispersion control in that case.

As for the objective lenses, apparently the 25x was designed

specifically for laser scanning applications, but I'd want to see a

side-by-side comparison with the 20x to be sure.

Craig

On Sun, Aug 23, 2009 at 2:03 PM, Bogdan Stoica<[hidden email]> wrote:

> Dear Lisa,

>

> Thank you for your kind response.

>  Let me adress the issue you raised in turn:

> -I did not get the chance to compare side by side the Olympus 25x vs. Leica

> 20x. I could only met with people that had rave reviews about the 25x so I

> would tend to five Olympus a slight advantage in this regard.

> -initially both systems had 4 NDDs but Leica had to leave only 2 to meet our

> budget. I saw people that having 4 NDDs coupled to 2 IR lasers and I was

> told me that even with one laser as in our case more NDDs could be

> beneficial for recording other channels such as intrinsic fluorescence etc.

> Again an extra point for Olympus.

> -You are right to point that for Leica features such as the AOBS as well as

> white laser should be unique advantages. The problem is that they would put

> the system outside our immediate budget although the fact that these

> upgrades are available in the future including as on-side upgrades for the

> AOBS justify an extra advantage for Leica.

> -Indeed, the need to do live cell imaging is directing us toward an inverted

> system.

> -the IR laser are the Maitai deepsee for olympus and the Chameleon Ultra II

> for the Leica.

>

> Bogdan

>

---------------------------------------------------------------------
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.
---------------------------------------------------------------------

[George McNamara]

Hi Bogdan,

In addition to the purchase price, you should obtain from the vendors the annual service contract(s) to calculate the total cost of ownership. Comparably equipped microscopes may have a factor of 2 difference in annual price, from different vendors. If your grant and project ends in two years, you may not care, but if you want to use the instrument as a confocal microscope in say year 7, 2x price difference in service contract is a big different in TCO.

The multiphoton laser service contract may be an additional cost direct from the company (ex. $15K/year for Coherent Chameleon II).

Leica offers an NDD4 module, though it is not attached to the microscope (I forget whether it is a liquid light guide or fiber optic). Our Leica NDD4 has blue, cyan, green and red filters, and additional filters are pricey compared to conventional microscope filter sets. Talk with Owen Schwartz about the Leica NDD4 implementation - he is the instigator for it.

Sincerely,

George
p.s. The New Zeiss LSM710NLO NDD# option is attached to the microscope.




At 12:11 PM 8/23/2009, you wrote:

Hi fellows,

I am also in the last stages of choosing a confocal/multiphoton system for
our laboratory. After seeing multiple demos i am thorn between an Olympus
FV1000MPE system and an Leica SP5 II. Our needs are to be able to do both
confocal images on fixed and live in vitro cells and live in vivo rodent
brain imaging. Both systems come with a UV, visible Argon four lines (488
etc), laser diode 560 and 633 and IR laser (Chameleon for Leica and Maitai
for Olympus), a complete set of Apo objectives as well as the famous 25x NA
1.05 from Olympus and the "equivalent" 20x from Leica. The microscopes are
inverted and the system also includes 2 NDD for Leica and 4 NDD in the case
for Olympus.
Fro what I could see the Leica software appeared more user friendly and I
would give their system a better feel for confocal imaging and manufacturers
potential upgrades- the system I am pricing does not include the AOBS
feature, "white laser" or resonant scanner but those could be added later.
On the other hand the Olympus seems a better choice for multiphoton imaging
and an easier system to tinker with. The tech support and managers for both
companies seemed outstanding to me here in the Washington DC-Baltimore area.
I found interesting that I was not able to find any info on "how much" such
a system should cost and after intense "negotiations" the quites range on my
case around the ~700k mark with Olympus a little bellow and Leica a little
above.
I was wondering if people who recently had to make a similar decision can
offer me some guidance both on the merits of their systems once they were
installed as well as on the price, support etc.
Thank you very much,

Bogdan

George McNamara, Ph.D.
Image Core Manager
Analytical Imaging Core Facility
University of Miami, Miller School of Medicine
Miami, FL 33010
[hidden email]
[hidden email]
305-243-8436 office
http://www.sylvester.org/AICF (Analytical Imaging Core Facility)
http://www.sylvester.org/AICF/pubspectra.zip (the entire 2000+ spectra .xlsx file is in the zip file)
http://home.earthlink.net/~geomcnamara

[Jerry (Gerald) Sedgewick]

A post-doctoral position is available at the Institute of Human
Virology, University of Maryland to join a laboratory studying the
mechanisms of entry and fusion of enveloped viruses. The primary focus
of the laboratory is on retrovirus entry (see /Miayuchi et al. Cell
2009/). Candidates with a Ph.D. degree in biomedical field or in
physics/chemistry who have at least 2 years of experience in live cell
confocal microscopy are encouraged to apply. The laboratory is equipped
with confocal and wide-field deconvolution imaging systems. IHV provides
excellent research support through core facilities and common equipment.
For more information, please visit the IHV web site: _www.ih.org
<http://www.ih.org/>_. Qualified applicants should submit a cover
letter, CV, and contact information for three references to Gregory
Melikian at [hidden email]
<mailto:[hidden email]>_.

Please note that Jerry Sedgewick is posting for Dr. Gregory Melikian, so
please do not reply to Jerry's email.  Reply, instead, to
[hidden email] <mailto:[hidden email]>._

--
Jerry (Gerald) Sedgewick
Core Facility Director, Biomedical Image Processing Lab (BIPL)
University of Minnesota, Department of Neuroscience
1-205 Hasselmo Hall
312 Church St. S.E.
Minneapolis, MN  55455
612-624-6607
[hidden email]
http://www.bipl.umn.edu
Author: "Scientific Imaging with Photoshop: Methods, Measurement and Output."

Rawlight.com (dba "Sedgewick Initiatives")
965 Cromwell Avenue
Saint Paul, MN  55114
651-788-2261
[hidden email]
http://www.quickphotoshop.com
http://www.rawlight.com
http://www.jerrysedgewick.com

[Aryeh Weiss]

Our Nikon rep has recommended to a C1 user to replace a failing 543nm
HeNe laser with an unpolarized 2mW laser. This C1 has no AOTF, and
combines three beams into a fiber which feeds the scanhead.

I was under the impression that using an unpolarized laser is a bad idea
because I recall that in an unpolarized HeNe, two orthogonal
polarization modes lase together, but they drift across the bandwidth of
the cavity as the laser warms up. This results in a slowly varying
change in the relative polariation of the beam over time. In a system
where reflections from dielectric surfaces can be dependent on
polarization, this should be a problem. If the fiber is polarization
preserving, it should be a big problem.

So -- does anyone out there know if my understanding is correct? Is
anyone using unpolarized HeNe lasers with no problem? I assume that
Nikon would not be offering that laser if there was such a problem
inherent to it, but still...

--aryeh
--
Aryeh Weiss
School of Engineering
Bar Ilan University
Ramat Gan 52900 Israel

Ph:  972-3-5317638
FAX: [phone]

[Sudipta Maiti]

Ususally the polarization changes pretty fast, certainly it does for my
unpolarized (randomly polarized) HeNe, so that I do not see any problems
in the image. I also
looked at the output fluorescence emission  at a 10 microsecond level
through a polarizer, and
saw no fluctuations. Of course, pixel residence times are somewhat smaller
than 10 microseconds.
Sudipta
  On Thu, 10 Sep 2009, Aryeh Weiss wrote:

> Our Nikon rep has recommended to a C1 user to replace a failing 543nm HeNe
> laser with an unpolarized 2mW laser. This C1 has no AOTF, and combines three
> beams into a fiber which feeds the scanhead.
>
> I was under the impression that using an unpolarized laser is a bad idea
> because I recall that in an unpolarized HeNe, two orthogonal polarization
> modes lase together, but they drift across the bandwidth of the cavity as the
> laser warms up. This results in a slowly varying change in the relative
> polariation of the beam over time. In a system where reflections from
> dielectric surfaces can be dependent on polarization, this should be a
> problem. If the fiber is polarization preserving, it should be a big problem.
>
> So -- does anyone out there know if my understanding is correct? Is anyone
> using unpolarized HeNe lasers with no problem? I assume that Nikon would not
> be offering that laser if there was such a problem inherent to it, but
> still...
>
> --aryeh
>

--
Sudipta Maiti
Dept. of Chemical Sciences
Tata Institute of Fundamental Research
Homi Bhabha Road, Colaba, Mumbai 400005, India
91-22-2278-2716
www.tifr.res.in/~biophotonics