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

Description

In this paper, we discussed how multi-dielectric coated coverslip can be designed to reach large optical field enhancement when working under proper illumination conditions and in total internal reflection. As objectivebased total internal reflection fluorescence microscopy (TIRF-M) is also based on total internal reflection illumination, 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.

Abstract

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