Abstract

The last decade has witnessed momentous advances in fluorescence microscopy.¹ The introduction of novel fluorescent markers,^2–4 together with the development of original microscopy techniques,^5–7 made it possible to study biomolecular interactions in living cells and to examine the structure and function of living tissues. The emergence of these innovative techniques had a. remarkable impact on all the life sciences. However, many biological and medical applications involve the detection of minute quantities of bionrolecules, and are limited by the signal weakness in common observation conditions. Here, we show that silver and gold-coated microscope slides can be used as mirror substrates to efficiently improve detection sensitivity when fluorescence microscopy is applied to micrometer-thick biological samples. We report a. fourfold enhancement of the fluorescence signal and a. noticeable strengthening of the image contrast, when mirror substrates are used with standard air microscope objectives. We demonstrate that metal-coated substrates provide the means to get sensitivity-enhanced fluorescence detection with dry optics, while keeping a. wide field observation and a. large depth of field. This is a. crucial advantage for automated and high-throughput applications to cell and tissue diagnostic analysis.

© 2007 SPIE