Just to keep the record straight, our Leica MP system has non-descanned
detectors in both transmission and epi directions - this was delivered
in 2000, ie 9 years ago. We were a fairly early customer for these. It
was (IMHO) a better implementation than the Bio-Rad one (less sensitive
to room light). Zeiss back then did have non-descanned detection but
only in transmitted direction - it worked quite well so long as your
sample wasn't impossibly thick.
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 http://www.guycox.net
______________________________________________
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]]
On Behalf Of Eric Scarfone
Sent: Thursday, 14 May 2009 12:46 AM
To: [log in to unmask]
Subject: Re: Recommendations for commercial multi-photon system purchase
Hi Lisa
Lucky you to purchase such a machine!!
The beauty with multiphoton excitation is that optical sectioning is
achieved by the limited volume within which photon density is sufficient
for 2 (or more) quasi simultaneous photon absorption events to take
place. For a given laser setting, the thickness within which this occurs
is only determined by the NA of your objective. ALL the fluorescence
coming from your sample is emitted by this NA determined plane. There is
no "out of focus" fluorescence. This is very different from Confocal.
To make full use of this one would like to collect as much of the
fluorescent photons emitted by the samples as possible. The most
effective way to do that is to use non-descanned external detectors both
in the reflected pathway and in the transmitted pathway.
To my knowledge this was implemented first by Warren Zipfel in Watt Webb
lab at Cornell and then commercialized by Bio-Rad on the last system
they put on the market before diseappearing at the end of the previous
century.... After a pretty long absorption/digestion process this was
implemented rather recently by Zeiss.
Others might have come up with similar solution but I do not know about
it.
I'd were you, for this level of price, I'd ask for an on site test with
your own samples. Do not rely on "in factory" demos!!
Cheers
Eric
Eric Scarfone, PhD, CNRS,
Center for Hearing and communication Research
Department of Clinical Neuroscience
Karolinska Institutet
Postal Address:
CFH, M1:02
Karolinska Hospital,
SE-171 76 Stockholm, Sweden
Work: +46 (0)8-517 79343,
Cell: +46 (0)70 888 2352
Fax: +46 (0)8-301876
email: [log in to unmask]
http://www.ki.se/cfh/
----- Original Message -----
From: "Cameron, Lisa" <[log in to unmask]>
Date: Wednesday, May 13, 2009 4:14 pm
Subject: Recommendations for commercial multi-photon system purchase
To: [log in to unmask]
> I have been investigating commercial multi-photon systems for
> awhile in order to
> purchase a system for my Institute's core microscopy facility. Our
> interest is
> to have the capability to do intravital imaging on an upright
> stand, but also be
> able to have facility users be able to put slides on and use the
> visible scanner
> and detectors. I realize this is a tall order for such versatility
> in one
> system, but since it is for a core (which needs to bring in
> revenue), I'm
> looking for the most flexible system. Does anyone have any
> suggestions about the
> most recent systems on the market? Or could you point out factors
> you think are
> the most important for making the decision on which company to go
> with? Please
> feel free to contact me off line.
>
> I have seen a demo of the Leica SP5 MP, Zeiss 710 NLO, Olympus MPE
> and Prairie's
> system.
>
> (BTW - my own experience is with widefield and confocal live-cell
> imaging, so I
> have not done 2-p myself, but have been learning about it for about
> a year)
>
> Thanks!
> - Lisa
>
> ---------------------------------------
> Lisa Cameron, Ph.D.
> Director of Confocal and Light Microscopy
> Dana Farber Cancer Institute
> 44 Binney St.; JF 215
> Boston, MA 02115
> Office phone: 617-582-8824
> Fax: 617-582-8750
> [log in to unmask]
>
>
>
> The information in this e-mail is intended only for the person to
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