Co is the initial concentration of the dye, while Cn is the concentration of the dye after dilution. Intensity of the images are found using ImageJ as well as MATLAB, there is hardly any difference in the grey values obtained from both. But the curve shows, otherwise, ie. increase in concentration with dilution, which is quite puzzling. Please help, Thank you
Mrs/Miss/Mr. Pore,
The topic is in the basic principles of teh spectroscopy an dthe environmental effects on the fluorophore intensity.
Please pay attention to this book:
http://www.springer.com/de/book/9780387312781
In simpler way it can be explained: As the concentration of solution increases there is increase in factor called internal quenching.
This internal quenching occurs due variety of processes like excited state reactions, energy transfer, complex-formation, collisional quenching, etc which get increased with increase in number of moles in the solution i.e. with in concentration of solution.
Hi Adity, I agree with Bharat. The most logical explanation is that you are diluting out quenching agents and thus increasing the fluorescence.
due to self quenching......upto certain concentration there is increase in fluorescence intensity with concentration , after that decrease in intensity with increasing concentration mainly due to collision among them.
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