Rapid immuno-SERS microscopy for tissue imaging with single-nanoparticle sensitivity.

Immuno-SERS microscopy is a novel imaging technique in nano-biophotonics, which employs antibodies labeled with SERS-active nanoparticles in conjunction with Raman microscopy. Rapid data acquisition is of central importance for screening large areas of tissue specimens. Here, we first discuss the role of SERS labels with single-particle sensitivity in immuno-SERS microscopy, in particular with respect to false-negative results. In combined single-particle experiments (SERS microscopy/dark-field microscopy/HR-SEM), we then demonstrate that small glass-coated clusters (dimers and trimers) of gold nanospheres exhibit the desired single-particle SERS sensitivity, even at acquisition times as short as 30 msec per pixel, while monomers do not. The proof-of-concept for rapid immuno-SERS microscopy with 30 msec acquisition time per pixel for selective imaging of the p53 family member p63 in prostate tissue sections is demonstrated.