Resonant multi-dielectric coverslip for enhanced total internal reflection fluorescence microscopy

Abstract

Multi-dielectric coated coverslip can be designed to reach large optical field enhancements when working under proper illumination conditions and in total internal reflection. In an objective-based total internal reflection fluorescence microscopy (TIRF-M), we propose to use the resulting large field enhancement supported in such coverslip to improve TIRF-M sensitivity by amplifying the collected fluorescence signal. We present here the optimization, realization and testing in TIRF-M of resonant coverslips designed to take into account the experimental constraints. The latter's are due to the inverted configuration of TIRF-M together with the use of a high numerical aperture objective. The challenge is therefore to find the best coating design compromising between the angular tolerance and the field enhancement. We will report here enhanced-TIRF-M imaging over model samples but the ultimate goal here is to be able to monitor and quantify dynamics of pathogens at the membrane of living cells.