Creating sensitive and reproducible substrates for surface-enhanced Raman spectroscopy (SERS) has been a challenge in recent years. While SERS offers significant benefits over traditional Raman spectroscopy, certain hindrances have limited their commercial use, especially in settings where low limits of detection are necessary. We studied a variety of laser-deposited silver microstructured SERS substrates with different morphology as a means to optimize analyte detection. We found that using a 405 nm laser to deposit lines of silver nanoparticles (AgNPS) from a 2 mM silver nitrate and sodium citrate solution offered not only the best enhancement, but also the most consistent and reproducible substrates. We also found that the probability of deposition by laser was wavelength dependent and that longer wavelengths were less likely to deposit than shorter wavelengths. This work offers a better understanding of the laser deposition process as well as how substrate shape and structure effect SERS signals.