Typical evanescent wave biosensors generate an electromagnetic wave at the sensor surface that penetrates 100–200 nm into the analysed medium. This has proven to be a highly sensitive tool to monitor refractive index changes in the close vicinity of the sensor surface. The sensitivity of such sensors can be enhanced significantly to monitor interactions caused by large micron scale objects such as bacterial and mammalian cells by increasing the penetration depth of the evanescent field. Recently, different formats of deep-probe optical waveguides including reverse waveguides (RW) based on low refractive index substrates (below 1.33) and metal-clad leaky waveguides (MCLW) have been developed for various sensing applications. These sensors are designed to maximize the overlap between the optical mode and the adlayer (superstrate layer) to be sensed. Increasing the penetration depth of the evanescent field opens up new perspectives for the detection of larger biological objects as it accommodates the majority of their body within the evanescent field. RWs use substrate materials with lower refractive index than that of the monitored superstrate layer (aqueous solution). In MCLWs, a thin metal layer is inserted between the substrate and the thicker waveguide layer. These sensor designs facilitate both increasing and tuning the penetration depth of the modes into the monitored aqueous solution and thereby significantly extend the range of possible application areas of optical waveguide sensors. The developed devices have been used for a range of biosensing applications, including the detection of bacteria, mammalian cells, organophosphorous pesticides and glucose using refractive index changes, absorbance and fluorescence monitoring. Integrating deep-probe sensors with an external electrical field or ultrasonic standing waves shortens analysis time significantly and reduces non-specific binding due to enhanced diffusion of analytes to the immobilized recognition receptors, and thus improves the detection limit by a few orders of magnitude.