Re: use of GFP and CFP reporters with a VantageSE

From: A Herman, Pathology & Microbiology (A.Herman@bristol.ac.uk)
Date: Tue Nov 05 2002 - 11:32:34 EST


Dear Bill,
 Thank you very much for your reply. The PI who asked me about CFP & GFP is
getting back to his collaborators re: other colour reporters for this grant
proposal.
  If the interest heightened we would explore adding another laser as you
suggested.
Thanks again, and just to add that I have found your website extremely
useful!

Yours,
Andy

--On Thursday, October 24, 2002 0:23 -0400 William Telford
<TelfordW@mail.nih.gov> wrote:

> Hi Andrew...
>
> CFP "likes" a violet or blue excitation source - krypton-ion 407 or 413
> nm violet, or argon-ion 457 nm blue lines all provide
> not-completely-optimal but decent excitation.  Unfortunately these lines
> can't be had from an Enterprise II.  488 nm won't work.
>
> The cheapest solution for CFP on the Vantage is to obtain a violet laser
> diode from Power Technologies or Coherent and set it up in the middle
> (third) laser position, where your UV beam is now.  These relatively
> low-power (10-20 mW) laser sources provide surprisingly good excitation
> of CFP.  You will need to set up some sort of mounting system - we've
> used a laser post mounting system from Newport.  You can set up the whole
> system for a good bit less than $10,000 - much less than a water-cooled
> gas laser.
>
> We have some sample data for violet laser diode excitation of CFP on our
> website at http://home.ncifcrf.gov/ccr/flowcore/index.htm (direct access
> at http://home.ncifcrf.gov/ccr/flowcore/VDL.htm).  If you are interested
> it trying this, we can send you more specific info.
>
> Good luck,
>
> Bill Telford
> NCI-NIH
>
>   At 12:11 PM 10/23/2002 +0100, Andrew Herman wrote:
>
>
> Dear Flow-ers,
>   I have a colleague who wishes to analyze and sort cells that bear GFP
> and CFP (cyan fluor. protein) reporter constructs.
> Has anyone had experience doing this type of analysis with a Vantage SE
> and an Enterprise II laser (6 color config). I realize that using a 488
> nm line is suboptimal for CFP, but is it feasible?
>
> Any info would be very much appreciated.
>
> Thanks,
>
> ----------------------
> Andrew Herman
> Department of Pathology & Microbiology
> University of Bristol
> University Walk
> Bristol  BS8 1TD
> Tel. +44 117 928 7511
> Fax. +44 117 928 7896
> A.Herman@bristol.ac.uk
>
>



----------------------
A Herman, Pathology & Microbiology
A.Herman@bristol.ac.uk



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