Surface-enhanced Raman scattering (SERS) probes are made by facile photochemical deposition of silver nanoparticles on a femtosecond (fs) laser ablated, D-shaped fiber. The structure and surface morphology of the probe are investigated by scanning electron microscopy. High-quality SERS signals are detected using Rhodamine 6G molecules via an in situ sensing mode. Experimental results show that the SERS signals increase with the increase of the length of fs laser ablated, D-shaped zone. Our D-shaped fiber SERS probe shows a feasible method for a large active area, high performance, and real-time and remote measurement of SERS signals in biochemical analysis.