I have amphiphilic nanoparticles that can encapsulate hydrophobic compounds. In order to observe it the cell by Fluorescence Microscopy, I decided to use DiO dye as a hydrophobic model. Before microscopy, I tested the fluorescence spectra of my nanoparticle interacted with DiO. I found a peak located at ~ 548 nm at excitation wavelength at 484 nm. 548 nm is different from 508 nm emission wavelength as normal DiO emission. Did anyone face to this issue? Is this peak showing me encapsulated DiO in hydrophobic environment?
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