New modeling of reflection interference contrast microscopy including polarization and numerical aperture effects: application to nanometric distance measurements and object profile reconstruction

Langmuir : the ACS Journal of Surfaces and Colloids
O TheodolyM-P Valignat

Abstract

We have developed a new and improved optical model of reflection interference contrast microscopy (RICM) to determine with a precision of a few nanometers the absolute thickness h of thin films on a flat surface in immersed conditions. The model takes into account multiple reflections between a planar surface and a multistratified object, finite aperture illumination (INA), and, for the first time, the polarization of light. RICM intensity I is typically oscillating with h. We introduce a new normalization procedure that uses the intensity extrema of the same oscillation order for both experimental and theoretical intensity values and permits us to avoid significant error in the absolute height determination, especially at high INA. We also show how the problem of solution degeneracy can be solved by taking pictures at two different INA values. The model is applied to filled polystyrene beads and giant unilamellar vesicles of radius 10-40 microm sitting on a glass substrate. The RICM profiles I(h) can be fitted for up to two to three oscillation orders, and extrema positions are correct for up to five to seven oscillation orders. The precision of the absolute distance and of the shape of objects near a substrate is about 5 nm i...Continue Reading

References

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Citations

Jul 4, 2012·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·Stoyan C RussevAtanas N Tzonev
Jul 22, 2010·Applied Optics·Jose C Contreras-NaranjoVictor M Ugaz
Mar 18, 2015·European Journal of Immunology·Alexandre BrodovitchPhilip Anton van der Merwe
Jul 1, 2020·Soft Matter·Lola CiapaAntoine Chateauminois
Dec 17, 2016·Nature Physics·Eva M SchmidDaniel A Fletcher
Feb 23, 2021·Biology of the Cell·Nicolas Garcia-SeydaOlivier Theodoly
Mar 10, 2012·Physical Review Letters·Jolet de RuiterFrieder Mugele
Aug 22, 2015·Physical Review Letters·Axel HuerreMarie-Caroline Jullien
Sep 4, 2020·Biophysical Journal·Laurene AounOlivier Theodoly

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