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(fwd) Re: BFP anyone?

Beat Ludin b_ware at COMPUSERVE.COM
Mon Aug 11 09:44:14 EST 1997


Alex Chang wrote:

>I'd like to know the filter set you are using for BFP (Quantum).
>
>I am co-expressiong both GFP and BFP, the GFP bled through the blue =

>filter quite a bit (using Zeiss, with filter#02).
>
>Alex

No wonder, as far as I remember, the Zeiss#02 has a longpass (LP) =

emission filter (that's why you see the GFP bleed as a GREEN signal). =

What you need is a bandpass (BP) emission filter to get rid of the GFP =

signal. Zeiss doesn't have a BP filter in their collection, but I'm sure =

they can get you one.
=2E
I'm using Chroma filters, mostly HQ. The specifications are (from the top=
 =

of my head, since I'm out of the lab):

ex       bs        em
405/?   420       460/50   for BFP   (brightest with mercury lamp)
390/20  420       460/50             (best S/N, especially if you have a =

xenon lamp)
360/?   420       460/50             (less bleed from GFP than the 405ex)=


480/30  500       525/40   for the GFP(S65T)

Good luck

           Beat

BTW, it seems to me, that EGFP is somewhat more UV-excitable than =

GFP(S65T) but I haven't had the chance to really measure it yet. Maybe =

somebody else can comment on this?





-----------------------------------------------------
Dr. Beat Ludin, FMI, Maulbeerstr 66, 4058 Basel, Switzerland
Tel. +41 61 697 6697 / FAX +41 61 697 3976
Internet:ludin at fmi.ch / Compuserve:100102,1527




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