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Thank you for the feedback. It certainly gives me/us something to think about.
Cheers,
Brian D Armstrong PhD
Assistant Research Professor
Light Microscopy Core
Beckman Research Institute
City of Hope
Dept of Neuroscience
1450 E Duarte Rd
Duarte, CA 91010
626-256-4673 x62872
http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Craig Brideau
Sent: Thursday, April 21, 2011 4:28 PM
To: [log in to unmask]
Subject: Re: Thorlabs confocals
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Actually we have the 85MHz repetition rate of the Octavius rather than the
1GHz version. We figured the individual pulse energy at 1 GHz was too weak.
85 MHz strikes a good balance for peak energy, and also allows for time for
any fluorophores to relax back to the ground state between pulses. I've
never actually tried the 1 GHz version though so your milage may vary.
There are good arguments on both sides of the "fast rep-rate and low peak
energy" vs "high peak energy and time to relax" sides of the table.
As to why we picked this rather than a MaiTai or Chameleon, we wanted to
excite multiple dyes simultaneously. The Octavius is so broad it tends to
excite everything all at once. From our preliminary mucking around we found
it will excite DAPI, Alexa 488, ROX, and a bunch of other dyes fairly well,
simultaneously. I'm currently working to improve the optics and do some
dispersion compensation to increase our peak energy. I'll let the list know
how it goes...
$50k buys you the Ti:Saph, but you still need your own pump laser I think.
We have a Verdi V8 driving ours, although the V5 also works I've been told.
It's not completely hands off, but I find it only needs minor adjustments on
roughly a weekly basis to keep it running in top form. The key thing is
that its output spectrum is stable and repeatable. An added bonus is it has
a very tall and narrow spike at 650 nm, so it has its own built-in
alignment/guide beam! We filter this out of the input beam so we can image
red dyes but it sure helps with getting the beam into the microscope
initially. I actually have a narrow band pass filter that only lets the 650
spike out so I can align without the full power of the laser flying around
the room.
Craig
On Thu, Apr 21, 2011 at 12:16 PM, Armstrong, Brian <[log in to unmask]>wrote:
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Craig, could you expound on why you chose this laser rather than a Coherent
> or Spectra Physics Ti:Sa laser (for example)?
>
> I see that the laser specs are; pulses are short (<6fs) and fast (1GHz) and
> less expensive (~$50k [I assume USD]).
>
> What will you use this laser for?
> (if you don't mind sharing)
>
> Thanks,
>
> Brian D Armstrong PhD
> Assistant Research Professor
> Light Microscopy Core
> Beckman Research Institute
> City of Hope
> Dept of Neuroscience
> 1450 E Duarte Rd
> Duarte, CA 91010
> 626-256-4673 x62872
>
>
> http://www.cityofhope.org/research/support/Light-Microscopy-Digital-Imaging/Pages/default.aspx
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:[log in to unmask]]
> On Behalf Of Craig Brideau
> Sent: Thursday, April 21, 2011 11:05 AM
> To: [log in to unmask]
> Subject: Re: Thorlabs confocals
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I have their Octavius laser stuck on a mostly conventional Nikon A1R.
> Peter
> Fendel, the fellow who actually designed the thing came out and personally
> installed it, then came for a repeat visit to help tweak it up and train me
> on the use of the laser. Whenever I've had any questions or even the
> slightest issue with the laser he has been extremely responsive. Our
> preliminary results with the laser are also very exciting... hope to have
> some nice images to brag about soon.
>
> Craig
>
>
>
> On Thu, Apr 21, 2011 at 9:49 AM, Vergara, Leoncio A. <[log in to unmask]
> >wrote:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Have you had a chance to arrange for an extended demo of the system? The
> > company could bring you a system so you can test it with your samples for
> a
> > few days... also, are you clear about the advantages and disadvantages of
> a
> > resonant scanner versus spinning disk and swept field systems?
> > A company with a smaller microscopy operation probably would not have the
> > service coverage of the traditional big microscope companies but you will
> > have a lot more personal contact with the development team.... and, yes,
> I
> > don't know anything about their systems but it sounds you will be a bit
> of a
> > beta tester... which may be part of the fun.
> >
> > Leoncio A. Vergara MD
> > Assistant Director
> > Center for Biomedical Engineering
> > Assistant Professor
> > Microbiology and Immunology
> > University of Texas Medical Branch
> > 409-750-2153 (cell)
> >
> > -----Original Message-----
> > From: Confocal Microscopy List [mailto:[log in to unmask]]
> > On Behalf Of Tim Feinstein
> > Sent: Thursday, April 21, 2011 10:37 AM
> > To: [log in to unmask]
> > Subject: Thorlabs confocals
> >
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > *****
> >
> > Hello all,
> >
> > Thorlabs had an interesting display of confocal systems last week, at
> > Experimental Biology in DC, that ranged from a multiphoton system to a
> > resonant-only confocal that they informally quoted at <$70k, including
> > lasers (!). The general theme seems to be stripped-down systems that do
> one
> > thing well; e.g. they are still considering whether or how to integrate a
> > FRAP function into their resonant device. I also wonder whether they
> have a
> > suitable support network in place to handle maintenance and upgrade
> issues.
> > As well, buying a new system with new software can make one feel like an
> > unpaid beta tester. We might include their resonant scanner in a search
> for
> > a live-cell dedicated scope, against spinning disc and swept field
> systems
> > from PE, Andor and Nikon (the Ti base is a must), but I am on the fence
> > about that.
> >
> > If anyone has experience with them, please feel free to contact me on
> line
> > or off.
> >
> > Thanks,
> >
> >
> > Tim
> >
> > Timothy Feinstein, PhD
> > Postdoctoral Fellow
> > Laboratory for GPCR Biology
> > Dept. of Pharmacology & Chemical Biology
> > University of Pittsburgh, School of Medicine
> > BST W1301, 200 Lothrop St.
> > Pittsburgh, PA 15261
> >
>
>
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