The detection of Raman signals inside living cells is a topic of great interest in the study of cell biology mechanisms and for diagnostic and therapeutic applications. This work presents the synthesis and characterization of flower-shaped gold nanoparticles and demonstrates their applicability as SERS-active tags for cellular spectral detection. The particles were synthesized by a facile, rapid new route that uses ascorbic acid as a reducing agent of gold salt. Two triarylmethane dyes which are widely used as biological stains, namely malachite green oxalate and basic fuchsin, were used as Raman-active molecules and the polymer mPEG-SH as capping material. The as-prepared SERS-active nanoparticles were tested on a human retinal pigment epithelial cell line and found to present a low level of cytotoxicity and high chemical stability together with SERS sensitivity down to picomolar particle concentrations.