. Significance: Hyperspectral imaging (HSI) provides rich spectral information for improved histopathological cancer detection. However, acquiring high-resolution HSI data for whole-slide imaging (WSI) can be time-consuming and requires a huge amount of storage space. Aim: WSI using a color camera can be achieved with fast speed, high image resolution, and excellent image quality due to the established techniques. We aim to develop an RGB-guided unsupervised hyperspectral super-resolution reconstruction method that is hypothesized to improve image quality while maintaining the spectral characteristics. Approach: High-resolution hyperspectral images of 32 histologic slides were obtained via automated WSI. High-resolution RGB histology images were registered to the hyperspectral images for RGB guidance. An unsupervised super-resolution network was trained to take the downsampled low-resolution hyperspectral patches (LR-HSI) and high-resolution RGB patches (HR-RGB) as inputs to reconstruct high-resolution hyperspectral patches (HR-HSI). Then, an Inception-based network was trained with the HR-RGB, original HR-HSI, and generated HR-HSI, respectively, for whole-slide histopathological cancer detection. Results: Our super-resolution reconstruction network generated high-resolution hyperspectral images with well-maintained spectral characteristics and improved image quality. Image classification using the original hyperspectral data outperformed RGB because of the extra spectral information. The generated hyperspectral image patches further improved the results. Conclusions: The proposed method potentially reduces image acquisition time, saves storage space without compromising image quality, and improves the image classification performance.

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Hyperspectral imaging (HSI) is an emerging modality for digital pathology. The purpose of this study is to develop an extended depth of field (EDOF) method for mosaic hyperspectral images acquired with a snapshot camera. EDOF is a technique for ensuring that an image is in focus at all points. A stack of mosaicked hyperspectral images of hematoxylin and eosin (H&E)-stained histologic slides were acquired at different positions along the z-axis and used to output a hyperspectral histologic image that was in-focus at every point. Three different methods were compared to achieve a fully focused image. We compared conventional patch-based methods to our proposed growth-based and band-based methods. The Brenner function was used to quantitatively measure the focus quality of each image measured. The results show that both of our proposed methods performed better qualitatively and quantitatively than the patch-based method, with the band-based method performing the best, as it leveraged dividing pixels into their proper wavelengths in addition to spatially, giving the algorithm better contrast to measure. In terms of speed, the band-based method was the fastest, followed by the patch-based method, with the growth-based method being the slowest. Our proposed extended depth of field hyperspectral imaging methods can have immediate applications in digital pathology, especially whole slide imaging, and other microscopic imaging.

. Corrections to the published article include values in Table 1 and provision of an omitted reference.