Abstract
This study demonstrates that spectral fingerprint patterns for a weakly scattering biological sample can be obtained reproducibly and reliably with tip-enhanced Raman spectroscopy (TERS) that correspond well with the conventional confocal Raman spectra collected for the bulk substance. These provided the basis for obtaining TERS images of individual self-assembled peptide nanotapes using an automated, objective procedure that correlate with the simultaneously obtained scanning tunneling microscopy (STM) images. TERS and STM images (64 × 64 pixels, 3 × 3 μm²) of peptide nanotapes are presented that rely on marker bands in the Raman fingerprint region. Full spectroscopic information in every pixel was obtained, allowing post-processing of data and identification of species of interest. Experimentally, the "gap-mode" TERS configuration was used with a solid metal (Ag) tip in feedback with a metal substrate (Au). Confocal Raman data of bulk nanotapes, TERS point measurements with longer acquisition time, atomic force microscopy images, and an infrared absorption spectrum of bulk nanotapes were recorded for comparison. It is shown that the unique combination of topographic and spectroscopic data that TERS imaging provides reveals differences between the STM and TERS images, for example, nanotapes that are only weakly visible in the STM images, a coverage of the surface with an unknown substance, and the identification of a patch as a protein assembly that could not be unambiguously assigned based on the STM image alone.
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