How to set up parameters when you collect luminescence emission(Rlu)...

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Ruisong Ye
Aug 7, 2020
protocol Protocol: Bioluminescence Resonance Energy Transfer (BRET) Assay for Determination of Molecular Interactions in Living Cells
How to set up parameters when you collect luminescence emission(Rlu) and fluorescence emission(YFP) simultaneously? (1)Both Rlu and YFP emission are measured in luminescence model, or (2) Rlu in luminescence model following by YFP in fluorescence model. which one is correct? Thanks!
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Laurence J. Miller Author Answered Aug 10, 2020

Mayo Clinic, United States

You should have a filter set to collect the emission of luciferase at luciferase emission wavelength (465 bandwidth 25) and the emission of YFP at the emission of YFP wavelength ( 535, bandwidth 25). So when you add coeleterazine h to excite the luciferase and the collect the emission of YFP.
Basically emission measured in luminescence model
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