Exciting with slide projector

Sat Jan 11 14:44:13 EST 1997

 Dear Christoph,

    Here is some information about my illumination techniques and apparatus. 
The yeast I am screening have an S65T version of GFP which has an 
excitation wavelength of 488nm and an emission wavelength of 512nm. For 
illumination I use a Kodak 4400 slide projector with an optical filter in 
the slide slot. There is nothing special about the slide projector that I 
know of; I think that any one with a decent bulb will work.  The filter 
is a 488DF22 narrow bandpass filter from Omega Optical (802/254-2690) 
matched to the excitation wavelength of the S65T gfp (488nm).  The filter 
I have is a 50mmX50mm square, the size of a slide. It  has low band 
blocking on it so that it will not transmit red light.  Since the 488nm blue
illuminating light is visible, a filter is needed to view the much less 
intense 512nm green fluorescence. I use two Kodak No.12 gelatin filters 
taped to a UV face sheild.  The UV face sheild is not needed but it is a
convinient way to hold the filters.
    THis is definitely not the most sensitive way to detect 
fluorescence.  I can detect much weaker fluorescence at the cell level 
using a fluorescence mircroscope or FACS, but I does work for my purposes.
     I hope this information will help.

Stephen Cronin
scronin at jeeves.ucsd.edu

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