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
