A method to fabricate novel surfaces for the enhancement of biomolecule detection is described. Superhydrophobic (via chemical modification) silicon nanowire (NW) surfaces with laser-ablated, hydrophilic (LAH) spots are used to improve the sensitivity of matrix-assisted laser-desorption ionization (MALDI) mass spectrometry. MALDI is currently one of the most widely used tools for identification of biomolecules. Advantages of MALDI include low detection limits, soft ionization and analysis of a wide range of biomolecules. However, the UV laser used to ionize/desorb the samples is typically only ∼0.100 mm in diameter, while the standard MALDI target spot is 2.5 mm in diameter. Thus, only a small fraction of the sample is desorbed/ionized with each shot of the laser. This work utilizes roughened, nanostructured surfaces with LAH spots to concentrate the samples down to a ∼0.250 mm diameter spot size, improving both efficiency of laser desorption/ionization and sensitivity of the instrument. In preliminary, fundamental studies, the NW surfaces were characterized in terms of wettability and pinning strength of the LAH sites. Results indicate the surfaces are superhydrophobic and the force of the pinning sites acting on a water droplet increases linearly as the LAH site diameter increases. Interfacial phenomena were also explored in terms of evaporation and crystallization processes. Acetonitrile exhibits a relatively high evaporation rate compared to water. When protein and/or matrix are added to the system, evaporation rate of acetonitrile drastically decreases. The correlation between qualitative observations of crystallization and MALDI signal were examined. Results suggest that crystallization is governed by kinetics and crystals that form relatively rapidly and efficiently are not ideal for MALDI conditions. Myoglobin samples at various concentrations were analyzed by MALDI on a standard MALDI plate, a Teflon-coated plate and on NW supports with LAH spots. Results indicate an average improvement in MALDI sensitivity over the standard MALDI plate for whole proteins by a factor of 5 and over the Teflon-coated plates by a factor of 1.5. Due to the ability for NW surfaces with LAH sites to efficiently confine and constrain protein/matrix samples, NW supports may be used for feasible automation of MALDI and coupling with separations techniques, such as capillary electrophoresis.