I immobilize fluorescent peptide on chitosan films and when I try to measure the thickness of the film with ellipsometry, the software says that I have a film with 2 nm or less. However, with chitosan films without immobilization of the peptide, I detect a thickness of 20 nm.
I already tried to change the model but it did not work. Is it possible that the fact of the peptide has fluorophores interferes with the measurement? Is it possible that the peptide is absorbing the wavelength?
Thank you so much!
Ellipsometry is very sensitive to Optical properties - like absorbance or fluorescence.
Do you have access to a profilometer or interferometric film thickness measurement ?
James E Hanson Unfortanelly, I don't think so. But the best excitation wavelength of my fluorophore is in the UV region, and the emission is practically the same that the wavelength of the incident light. Do you think that is the problem?
If wavelength is the same as emission you are tuned to a transition that might interfere. Is the wavelength variable?
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