[Fluorescent-proteins] Question about fluorescence_can you post it
on the forum?
(by l.depiscopo08 from imperial.ac.uk)
Tue Aug 14 06:01:36 EST 2012
Dear scientific community,
I am a PhD student form Imperial College London, I need help with a fluorescence-related issue.
I am spotting a solution of quinine on aldehyde-coated glass slides. The quinine is solubilised in 0.1M sulfuric acid.
I am scanning the slide with a fluorescence scanner. The laser is set at 532nm and the (emission) filter is set at 575nm. Based on the absorption/emission spectrum of quinine, there should be no absorption at all above 500nm, and therefore I assumed that no fluorescence should be emitted/visualised. For the propose of our experiment, we have to prove that NO SIGNAL is detected at these settings.
However, when I scan the slide with these parameters, I record a very strong signal from the quinine spots. Why is that? Can anyone help?
The options are: 1) sulfuric acid reacts with the aldehydes on the slides and then somehow modifies the fluorescence properties of the quinine molecules; 2) I (and my colleagues here) don't understand fluorescence of quinine properly; 3) our scanner has problems, which I am tempted to exclude because it's just undergone maintenance.
Thanks very much
Lorenzo d'Episcopo +44(0)7896931761
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