khatipovNO at NOuchicago.edu
Fri Sep 28 16:35:36 EST 2001
As in most instruments of that kind (I have no experience with your
particular fluorometer), everything should be quite self explanatory from
browsing through the menus of the software that runs the instrument. You can
basically select the type of plate you are using, set the gain level, define
excitation and emission wavelengths, define which wells are controls, and
which are unknown, etc. You will have to determine the sensitivity of your
particular assay anyway. Sensitivity of the instrument should be given in
the manual. If not, it does not really matter much, because in a certain
sense designers of the instrument should have made it to have most optimal
sensitivity range for our everyday applications. On the other hand, you can
increase readout by changing the gain level, although keep in mind that it
also means increasing the noise. Just play around with the instrument and se
what it does. You might want to, e.g., take several serial dilutions of
purified GFP, read fluorescence under conditions (buffer, volume, etc) of
your experiment, and knowing the quantity of GFP in test solution, plot
fluorescence versus protein quantity to determine sensitivity range.
"yy" <yijst1 at pitt.edu> wrote in message news:3BB24039.5CECEBD4 at pitt.edu...
> hi all,
> we have a LS50B flurometer with well plate reader in the dept. i am
> planning to measure some GFP on that. but there are not much information
> in the manual, such as the low limit of the detection, what kind of
> plate i should use, etc.
> Can any one with experience with this machine give me some advice? the
> more the better.
> thanks ahead!
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