Chemiluminescence and the PhosphorImager?

Tracy Aquilla aquilla at salus.med.uvm.edu
Mon Nov 21 10:52:52 EST 1994


In Article <3als1u$fpl at lastactionhero.rs.itd.umich.edu>, "RNeubig at umich.edu"
<Rick Neubig> wrote:
>> Can one detect chemiluminescence with a PhosphorImager? 
>
>
>The most common PhosphorImager (at least around our place) is 
>the Molecular Dynamics. That one will not read Chemiluminescence
>though they have a separate machine that will read fluorescence.
>
>The Bio-Rad phosphorimager uses a somewhat different technology
>and is able to read both chemiluminescence and radioactivity
>though you need a separate screen for each.

Bio-Rad's GS-250 Molecular Imager can detect chemiluminescence quite well,
and you don't necessarily need separate screens to detect both radioactivity
and chemiluminescence. Bio-Rad's imager has three different types of screens
available, two optimized for radioactivity (BI and HS), and another
optimized for chemiluminescence (CH). The machine I have access to is used
by many different labs, most of whom buy their own screens and loading docks
for doing the exposures. It seems many labs around here buy the screens
optimized for use with 32P (BI screen), but I could not find anyone with a
screen optimized for chemiluminescence (CH screen). Bio-Rad's tech support
said the BI screen should work OK for detecting chemiluminescence, so I
tried it. This worked very well for me. Using Amersham's ECL and x-ray film,
I was getting over-exposure of bands on Westerns after just a 1 second
exposure, so I tried exposing these blots to the Bio-Rad BI screen for
fifteen minutes instead. Using the BI screen, I got a linear response from
bands that had appeared all black using the ECL and x-ray film. Thus, I
found the BI screen to be better than x-ray film for my westerns, since the
linear range of the medium is so much better, and the time needed for
exposure was not significantly different for me. This system makes
quantitation of nucleic acids and proteins a breeze. Of course, using the CH
screen for chemiluminescence would be better, since it would be even more
sensitivethan the BI screen.

Tracy Aquilla, Ph.D.
Molecular Physiology and Biophysics
University of Vermont
aquilla at salus.med.uvm.edu



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