Ultra-sensitive, rapid detection of dried bloodstains by surface enhanced Raman scattering on Ag substrates

A simple water extraction and transfer procedure is found to result in reproducible and highly sensitive 785 nm excited SERS spectra of 24 h dried bloodstains on Ag nanoparticle substrates. This protocol allows confirmatory detection and identification of dried stains of blood that have been diluted by up to 10⁵ in water on Ag substrates. While previous SERS results demonstrated similar performance on Au substrates when a 50% acetic acid extraction and transfer procedure was used, the water/Ag methodology avoids any potential DNA damage when the sample size is extremely small (≤∼1 μL) due to low pH exposure. The water only procedure is not effective on Au SERS substrates. This metal substrate difference results from the efficient red blood cell lysis and hemoglobin denaturation effects of the Ag nanoparticle surfaces as compare to that of Au nanoparticles. Consequently, the 50% acetic acid exposure is required for the acquisition of 785 nm SERS spectra of dried bloodstains on Au substrates.