I need to measure fluorescence excitation-emission spectra with TECAN Infinite M1000 Pro. I use the i-control 1.11 software to control the plate reader. I need to measure the whole range of excitation (230-850 nm) and emission (280-850 nm). The software offers a protocol for 3D spectra scan. However, by setting ex=230-850 and em=280-850, also unwanted combinations of wavelegths would be measured:
a) ex>em results in nonsense data and doubles the measuring time for each well. I also get nonzero values with this combination of ex-em settings and that is weird.
b) ex=em gives too high signal (oversaturation) and could damage the detector.
Is there any way how to set the emission to be always dependent from the excitation e.g. em=ex+10 nm ? Has anyone used the this software-hardware for the full 3D ex-em spectra?
Jp Wu - I know, that is because I want to avoid measuring ex=em. Any idea how to do that?
in fact, there is no point to measure ex>em as in principle em is always avove ex. hence in practice, you can start with min ex and set em from ex plus 50 up to max em. then ex with increment of 10 followed by em from ex+50 up to max em and so on. if your system is good enough to minmise the impact of ex on em, you can gradually reduce +50 to e.g. +10, etc. by doing this, you will get a series of fluresence spec of em against ex. you can display it in 2D by using, e.g. matlab, if you know how to use matlab. you will get a nice 2D display and can obtain the optimised ex and em of your materials if you need.
Jp Wu - I know all of this. The problem is that the software apparently does not give me the option to set em=ex+10. The only option that I found is to set fixed full range of ex 230-850nm and em 280-850 nm. That, however, results in measuring also ex>em and ex=em.
My question was whether anyone is using the same software and knows how to set the ex and em wavelengths to obtain sensible dataset.
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