I am looking at single molecules embedded in a polymer matrix, but there is a significant amount of background contamination in the polymer matrix that will Experimentally, what is the best way to try and reduce the concentrations of the contaminants?
I have already checked to see if it is the coverslip, tried to precipitate the polymer, have ran the polymer through a silica gel column, and have tried different polymers from different suppliers.
Try size exclusin chromatography (SEC); it is widely used for polymer purification.
I have used polymer films for these type of experiments and in general the background was reasonably low at excitation wavelenght in the green but it was much worse when the excitation was 405nm. The best results were using the highest purity polymer available commercially. What polymer are you using?
Jose--we're currently using either PS or PMMA. I've tried both polymer source and sigma. We've been thinking about making things in house if we can't get either of those to work.
Philipp--Our excitation wavelength is stuck at 473nm, unfortunately. What is considered high intensity for these types of applications? I have access to a ~70 mW 375nm laser that I have used to photobleach the polymer with some success.
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