I want to do a high throughput luminescence reading using 96 well plates based on firefly luciferase system. However I can only find a microplate reader equipped with fluorescence detector. There is no luminescence detector module available for this reader.
Can I use the fluorescence optics to measure luminescent reactions? Since both fluorescence and luminescence detection are just counting photons, except fluorescence needs light source for excitation.
Is it reliable? I guess because of the filter, I would lose some light signals.
Hi, guys. thanks very much for your input.
I have asked a specialist from Tecan. I showed a link to an BioTek specialist's note online. http://www.biotek.com/resources/articles/measure-luminescence-reactions.html
This what he said in the email.
"It is possible to detect light from luminescence with electronics and optics that are primarily intended for fluorescence but they lack sensitivity and signal dynamic range. The detector that Biotek is using for their luminescence “hack” is an analog photomultiplier tube (PMT) that relies upon different gain settings to place it in a state where the wells can be measured without under or over amplifying the signal. Tecan readers equipped with luminescence detection, as well as other luminometers, all use Photon Counting PMT detectors for luminescence. Photon counting detectors do not require adjustable gain settings. Gain is fixed at the factory and is never changed. Photon counting detectors have very low noise for very high signal to noise ratios(sensitivity) and a gigantic dynamic range that is upwards of 109. This makes them ideal for luminescence assays."
Hi , sorry to say the information in the above answer is not good. for a luminescence assay you are not measuring a specific wavelength of light but total light , there is no excitation source either as per fluorescence.
The correct method for this type of assay is : in the filter wheel for excitation you have a black plug which blocks light transmission from the excitation source. in the emission filter wheel you remove a filter so that there is just a hole and you get total light transmission.
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