Emerging clinical interest in combining standard white light endoscopy with targeted near-infrared (NIR) fluorescent contrast agents for improved early cancer detection has created demand for multimodal imaging endoscopes. We used two spectrally resolving detector arrays (SRDAs) to realize a bimodal endoscope capable of simultaneous reflectance-based imaging in the visible spectral region and multiplexed fluorescencebased imaging in the NIR. The visible SRDA was composed of 16 spectral bands, with peak wavelengths in the range of 463 to 648 nm and full-width at half-maximum (FWHM) between 9 and 26 nm. The NIR SRDA was composed of 25 spectral bands, with peak wavelengths in the range 659 to 891 nm and FWHM 7 to 15 nm. The spectral endoscope design was based on a "babyscope" model using a commercially available imaging fiber bundle. We developed a spectral transmission model to select optical components and provide reference endmembers for linear spectral unmixing of the recorded image data. The technical characterization of the spectral endoscope is presented, including evaluation of the angular field-of-view, barrel distortion, spatial resolution and spectral fidelity, which showed encouraging performance. An agarose phantom containing oxygenated and deoxygenated blood with three fluorescent dyes was then imaged. After spectral unmixing, the different chemical components of the phantom could be successfully identified via majority decision with high signal-to-background ratio (>3). Imaging performance was further assessed in an ex vivo porcine esophagus model. Our preliminary imaging results demonstrate the capability to simultaneously resolve multiple biological components using a compact spectral endoscopy system.
The ability to resolve multiple fluorescent emissions from different biological targets in video rate applications, such as endoscopy and intraoperative imaging, has traditionally been limited by the use of filter‐based imaging systems. Hyperspectral imaging (HSI) facilitates the detection of both spatial and spectral information in a single data acquisition, however, instrumentation for HSI is typically complex, bulky and expensive. We sought to overcome these limitations using a novel robust and low cost HSI camera based on a spectrally resolved detector array (SRDA). We integrated this HSI camera into a wide‐field reflectance‐based imaging system operating in the near‐infrared range to assess the suitability for in vivo imaging of exogenous fluorescent contrast agents. Using this fluorescence HSI (fHSI) system, we were able to accurately resolve the presence and concentration of at least 7 fluorescent dyes in solution. We also demonstrate high spectral unmixing precision, signal linearity with dye concentration and at depth in tissue mimicking phantoms, and delineate 4 fluorescent dyes in vivo. Our approach, including statistical background removal, could be directly generalised to broader spectral ranges, for example, to resolve tissue reflectance or autofluorescence and in future be tailored to video rate applications requiring snapshot HSI data acquisition.
Evaluation of illumination system uniformity for wide-field biomedical hyperspectral imaging View the table of contents for this issue, or go to the journal homepage for more 2017 J. Opt. 19 045301
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