Simultaneous refractive index and thickness measurement with the transmission interferometric adsorption sensor

Refractive index and thickness of the adlayer are determined simultaneously using the transmission interferometric adsorption sensor (TInAS). Optical biosensors, where both refractive index and thickness of a homogeneous adlayer (thus the adsorbed mass) are determined simultaneously, so-called model-free biosensors, are important tools to investigate the adsorbed mass of biomolecules with unknown conformation. Our proposed calculation method enables model-free biosensing from a single spectrum acquired by a simple TInAS setup, namely using information of peak/dip positions as well as peak/dip intensities. The feasibility of this method was experimentally tested by adsorbing polyelectrolyte multilayer as well as biomolecules. To validate the new method also for the more intricate heterogeneous adlayer, the apparent refractive index and thickness were assessed theoretically by simulating a selection of different adsorbate configurations with the multiple multipole program (MMP). We found that a lateral inhomogeneity of the adsorbate (e.g. islands or adsorbed colloids) results in correct thickness and in reduced refractive index averaged in proportion to their density while vertically inhomogeneous density caused more complex responses. However, the apparent mass was always correct. Measurement errors can lead to significant errors in the apparent refractive index, particularly when the adlayer is very thin (<5 nm). This model-free TInAS technique would be useful not only for the measurement of adsorbed mass but also for the conformational analysis of the adsorbed molecules.