Reduction of Spectral Overlap in Spectral Surface-Enhanced Raman Spectroscopy Imaging Using a Dove Prism

The ability to combine microscopy and spectroscopy is beneficial for directly monitoring physical and biological processes. Spectral imaging approaches, where a transmission diffraction grating is placed near an imaging sensor to collect both the spatial image and spectrum for each object in the field of view, provide a relatively simple method to simultaneously collect images and spectroscopic responses on the same sensor. Initially demonstrated with fluorescence spectroscopy, the use of spectral imaging in Raman spectroscopy and surface-enhanced Raman spectroscopy (SERS) can provide a vibrational spectrum containing molecularly specific information that can inform on chemical changes. However, a major complication to this approach is the spectral overlap that occurs when objects are spaced closely together horizontally. In this work, we add a dove prism to a spectral imaging instrument developed for SERS imaging, enabling rotation of the collected SERS image and dispersed spectrum onto the imaging complementary metal-oxide semiconductor (CMOS) sensor. We demonstrate that this effectively reduces spectral overlap for emitters with clear separation between them and emitters with slightly overlapping point spread functions thereby facilitating collection of unambiguous spectra from each emitter.