In this work, a novel nanostructure of gold nanosheets (NSs)-decorated TiO2 nanocolumns (NCs) coated on carbon fiber (Au NSs-TiO2 NCs/CF) is designed and fabricated. The structure and morphology can be adjusted from Tanghulu-like to sausage-shaped with the final concentration of chloroauric ions in the reaction solution. Au NSs-TiO2 NCs/CF have been proven to be an excellent SERS substrate with a high analytical enhancement factor, good uniformity, and great stability. Most importantly, SERS experiments can be carried out on one strand of Au NSs-TiO2 NCs/CF with the aid of a Raman confocal microscope. These Tanghulu-like and sausage-shaped Au NSs-TiO2 NCs/CF have great potential in SERS analysis of trace samples.Graphical abstract: