Rapid and sensitive detection of chemical and biological analytes becomes increasingly important in areas such as medical diagnostics, food control and environmental monitoring. Optical biosensors based on surface plasmon resonance (SPR) and optical waveguide spectroscopy have been extensively pushed forward in these fields. In this book, we combine long range surface plasmons (LRSPs), surface plasmon-enhanced fluorescence spectroscopy (SPFS) and optical waveguide spectroscopy with hydrogel thin film for highly sensitive detection of molecular analytes. LRSPs are special surface plasmon waves propagating along thin metal films with orders of magnitude higher electromagnetic field intensity and lower damping than conventional SPs. Therefore, their excitation on the sensor surface provides higher fluorescence signal for molecules detection. The hydrogel thin film attached on a sensor surface served both as a binding matrix and as an optical waveguide for probing molecules interactions. The higher binding capacity of hydrogel and low damping of optical waveguide provide higher sensitivity for molecules detection as compared to regular SPR.