Abstract
The intrinsic near-infrared photoluminescence (fluorescence) of single-walled carbon nanotubes exhibits unique photostability, narrow bandwidth, penetration through biological media, environmental sensitivity, and both chromatic variety and range. Biomedical applications exploiting this large family of fluorophores will require the spectral and spatial resolution of individual (n,m) nanotube species’ fluorescence and its modulation within live cells and tissues, which is not possible with current microscopy methods. We present a wide-field hyperspectral approach to spatially delineate and spectroscopically measure single nanotube fluorescence in living systems. This approach resolved up to 17 distinct (n,m) species (chiralities) with single nanotube spatial resolution in live mammalian cells, murine tissues ex vivo, and zebrafish endothelium in vivo. We anticipate that this approach will facilitate multiplexed nanotube imaging in biomedical applications while enabling deep-tissue optical penetration, and single-molecule resolution in vivo.
Highlights
Spectral imaging is a powerful tool for detection, validation, separation, and quantification in applications ranging from mineral assessment of geological satellite images[14] to semiconductor material characterization[15]
12 distinct fluorescent nanotube species were simultaneously detected in a 500 nm imaging window
We constructed a near-infrared hyperspectral microscope by incorporating a volume Bragg grating between the emission port of a standard inverted fluorescence microscope and the nIR camera (Fig. 1a) to obtain the spectral properties of the spatially-resolved emitted light
Summary
Spectral imaging is a powerful tool for detection, validation, separation, and quantification in applications ranging from mineral assessment of geological satellite images[14] to semiconductor material characterization[15]. We developed a wide-field near-infrared hyperspectral microscopy approach to spatially observe the fluorescence and spectral heterogeneity from single nanotubes in complex environments, including live cells and tissues. Exploiting the narrow spectral bandwidth (full width at half maximum, FWHM) of nanotubes, we resolved 17 distinct chiralities of individual nanotubes on a surface using a single excitation laser.
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