Detection and identification of biomacromolecules is of critical importance in many fields ranging from biotechnology to medicine. Surface-enhanced Raman scattering (SERS) is an emerging technique for the label-free detection and identification of biological molecules and structures with its fingerprinting properties and high sensitivity. However, there are a number of obstacles for its applications for biological macromolecules due to their complexity. In this report, manipulation of microscopic processes in play during the drying of a sessile droplet as a tool to influence the nanoparticle-macromolecule packing, which has dramatic effect on SERS performance, before the SERS acquisition is demonstrated. A process known as the coffee ring phenomenon jams all particles and molecular species to the edges of the droplet during drying. This uncontrolled process has dramatic effects on a SERS experiment, using colloidal metal nanoparticles as substrates, by sweeping everything to the edges and influencing the packing of nanoparticles in the droplet area. A plastic tip was dipped into a drying sample droplet to influence the uncontrolled piling up. A negatively-charged protein, BSA, a positively-charged protein, cytochrom c, and a 20-base long oligonucleotide, were used as model biomacromolecules in this study. While a minimum of one order of magnitude lower concentration improvement in detection limit was observed with negatively-charged biomacromolecules, no significant improvement was observed with positively-charged ones compared to a sample droplet left on the surface without any interference. With the demonstrated approach, picomolar-level biomolecular detection using SERS is possible.© (2011) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.