Multimodal hyperspectral optical microscopy

• Published in: Chemical physics 2017-11, Vol.498-499, p.25-32
• Main Authors: Novikova, Irina V., Smallwood, Chuck R., Gong, Yu, Hu, Dehong, Hendricks, Leif, Evans, James E., Bhattarai, Ashish, Hess, Wayne P., El-Khoury, Patrick Z.
• Format: Article
• Language: English
• Subjects: BASIC BIOLOGICAL SCIENCES; Bioimaging; Environmental Molecular Sciences Laboratory; Hyperspectral optical microscopy; Multimodal optical microscopy; OTHER INSTRUMENTATION; Spectral imaging microscopy
• ISSN: 0301-0104
• DOI: 10.1016/j.chemphys.2017.08.011

[Display omitted] •A convenient approach to multimodal hyperspectral optical microscopy is described.•Hyperspectral fluorescence measurements are used to calibrate our setup.•Different applications of hyperspectral dark field microscopy are presented.•Different applications of hyperspectral optical absorption microscopy are described. We describe a unique approach to hyperspectral optical microscopy, herein achieved by coupling a hyperspectral imager to various optical microscopes. Hyperspectral fluorescence micrographs of isolated fluorescent beads are first employed to ensure spectral calibration of our detector and to gauge the attainable spatial resolution of our measurements. Different science applications of our instrument are then described. Spatially over-sampled absorption spectroscopy of a single lipid (18:1 Liss Rhod PE) layer reveals that optical densities on the order of 10−3 can be resolved by spatially averaging the recorded optical signatures. This is followed by three applications in the general areas of plasmonics and bioimaging. Notably, we deploy hyperspectral absorption microscopy to identify and image pigments within a simple biological system, namely, a single live Tisochrysis lutea cell. Overall, this work paves the way for multimodal spectral imaging measurements spanning the realms of several scientific disciplines.