An analytical platform suitable for trace detection using a small volume of analyte is pertinent to the field of toxin detection and criminology. Plasmonic nanostructures provide surface-enhanced Raman scattering (SERS) that can potentially achieve trace toxins and/or molecules detection. However, the detection of highly diluted, small volume samples remains a challenge. Here, we fabricate a superhydrophobic SERS platform by assembling Ag nanocubes that support strong surface plasmon and chemical functionalization for trace detection with sample volume of just 1 μL. Our strategy integrates the intense electromagnetic field confinement generated by Ag nanocubes with a superhydrophobic surface capable of analyte concentration to lower the molecular detection limit. Single crystalline Ag nanocubes are assembled using the Langmuir-Blodgett technique to create surface roughness. To create a stable superhydrophobic SERS platform, an additional 25 nm Ag coating is evaporated over the Ag nanocubes to "weld" the Ag nanocubes onto the substrate followed by chemical functionalization with perfluorodecanethiol. The resulting substrate has an advancing contact angle of 169° ± 5°. Our superhydrophobic platform confines analyte molecules within a small area and prevents the random spreading of molecules. An analyte concentrating factor of 14-fold is attained, as compared to a hydrophilic surface. Consequently, the detection limit of our superhydrophobic SERS substrate reaches 10(-16) M (100 aM) for rhodamine 6G using 1 μL analyte solutions. An analytical SERS enhancement factor of 10(11) is achieved. Our protocol is a general method that provides a simple, cost-effective approach to develop a stable and uniform superhydrophobic SERS platform for trace molecular sensing.