Posted by
Guy Cox on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Re-Recommendations-for-commercial-multi-photon-system-purchase-tp3499069p3516814.html
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 +61 2 9351 3176 Fax +61 2 9351 7682
Mobile 0413 281 861
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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
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Sylvie Le Guyader
Dept of Biosciences and Nutrition
Karolinska Institutet
Novum
14157 Huddinge
Sweden
+46 (0)8 608 9240
>
> -----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
>
>
>
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