Spectral effects and enhancement quantification in healthy human saliva with surface‐enhanced Raman spectroscopy using silver nanopillar substrates

Objectives Raman spectroscopy as a diagnostic tool for biofluid applications is limited by low inelastic scattering contributions compared to the fluorescence background from biomolecules. Surface‐enhanced Raman spectroscopy (SERS) can increase Raman scattering signals, thereby offering the potential to reduce imaging times. We aimed to evaluate the enhancement related to the plasmonic effect and quantify the improvements in terms of spectral quality associated with SERS measurements in human saliva. Methods Dried human saliva was characterized using spontaneous Raman spectroscopy and SERS. A fabrication protocol was implemented leading to the production of silver (Ag) nanopillar substrates by glancing angle deposition. Two different imaging systems were used to interrogate saliva from 161 healthy donors: a custom single‐point macroscopic system and a Raman micro‐spectroscopy instrument. Quantitative metrics were established to compare spontaneous RS and SERS measurements: the Raman spectroscopy quality factor (QF), the photonic count rate (PR), the signal‐to‐background ratio (SBR). Results SERS measurements acquired with an excitation energy four times smaller than with spontaneous RS resulted in improved QF, PR values an order of magnitude larger and a SBR twice as large. The SERS enhancement reached 100×, depending on which Raman bands were considered. Conclusions Single‐point measurement of dried saliva with silver nanopillars substrates led to reproducible SERS measurements, paving the way to real‐time tools of diagnosis in human biofluids.