While fluorescence image-guided surgery offers improved treatment outcomes for patients with cancer by permitting theidentification of tumors during resection, it has been plagued by slow translation into clinical practice due to the lengthyand costly approval process for fluorescent molecular markers. Label-free approaches to image-guided surgery providean alternative by discriminating between cancerous and noncancerous tissue based on differences in spectral reflectanceand autofluorescence between the tumor microenvironment and the surrounding anatomy. Unfortunately, state-of-the-arthyperspectral imaging systems capable of monitoring spectral differences across the entire surgical site utilize complexoptomechanical architectures that contribute to low image resolutions, low frame rates, and co-registration error thatcannot be calibrated, making these instruments impractical during demanding surgical workflows. To provide label-freesurgical guidance while addressing limitations with existing systems, we have developed a single-chip snapshothyperspectral imaging system that provides 27 spectral bands from ~450 nm to ~750 nm. By monolithically integrating astacked photodiode image sensor with pixelated interference filters, we have produced a highly compact imaging systemthat achieves a resolution of 1252-by-852 pixels at a rate of 17.2 frames per second while avoiding co-registration error.The system provides a signal-to-noise ratio of ~55 dB and a dynamic range of ~62 dB, and it can enable spectraldiscrimination under standard broadband surgical light sources. Preclinical images of human prostate tumor implants ina murine model have been examined and presented to demonstrate that the imaging system can differentiate betweencancerous and noncancerous tissue and can discriminate between distinct cancer types.
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