Breast cancer is the most common cancer among women worldwide, with an estimated 1.7 million cases and 522,000 deaths in 2012. Breast cancer is diagnosed by histopathological examination of breast biopsy material but this is subjective and relies on morphological changes in the tissue. Raman spectroscopy uses incident radiation to induce vibrations in the molecules of a sample and the scattered radiation can be used to characterise the sample. This technique is rapid and non-destructive and is sensitive to subtle biochemical changes occurring at the molecular level. This allows spectral variations corresponding to disease onset to be detected. The aim of this work was to use Raman spectroscopy to discriminate between benign lesions (fibrocystic, fibroadenoma, intraductal papilloma) and cancer (invasive ductal carcinoma and lobular carcinoma) using formalin fixed paraffin preserved (FFPP) tissue. Haematoxylin and Eosin stained sections from the patient biopsies were marked by a pathologist. Raman maps were recorded from parallel unstained tissue sections. Immunohistochemical staining for estrogen receptor (ER) and human epidermal growth factor receptor 2 ( HER2/neu) was performed on a further set of parallel sections. Both benign and cancer cases were positive for ER while only the cancer cases were positive for HER2. Significant spectral differences were observed between the benign and cancer cases and the benign cases could be differentiated from the cancer cases with good sensitivity and specificity. This study has shown the potential of Raman spectroscopy as an aid to histopathological diagnosis of breast cancer, in particular in the discrimination between benign and malignant tumours.

Histopathology and immunohistology remain the gold standard for breast cancer diagnostic. Yet, these approaches do not usually provide a sufficiently detailed characterization of the pathology. The purpose of this work is to demonstrate for the first time that elemental analysis and Fourier transform infrared spectroscopy microscopic examination of breast tissue sections can be merged into one dataset to provide a single set of markers based on both organic molecules and inorganic trace elements. For illustrating the method, 6 mammary tissue sections were used. Fourier transform infrared (FTIR) spectroscopy images reported a fingerprint of the organic molecules present in the tissue section and laser ablation elemental analysis (LA-ICP-MS) images brought inorganic element profiles. The 6 tissue sections provided 31 10 6 and 150,000 spectra for FTIR and LA-ICP-MS spectra respectively. The results bring the proof of concept that breast tissue can be analyzed simultaneously by FTIR spectroscopy and laser ablation elemental analysis (LA-ICP-MS) to provide in both case reasonably high resolution images. We show how to bring the images obtained by the two methods to a same spatial resolution and how to use image registration to analyze the data originating from both techniques as one block of data. We finally demonstrates the elemental analysis is orthogonal to all FTIR markers as no significant correlation is found between FTIR and LA-ICP-MS data. Combining FTIR and LA-ICP-MS imaging becomes possible, providing two orthogonal methods which can bring an unprecedented diversity of information on the tissue. This opens a new avenue of tissue section analyses providing unprecedented diagnostic potential.

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The spectral imaging technique has been shown to provide more discriminative information than the RGB images and has been proposed for a range of problems. There are many studies demonstrating its potential for the analysis of histopathology images for abnormality detection but there have been discrepancies among previous studies as well. Many multispectral based methods have been proposed for histopathology images but the significance of the use of whole multispectral cube versus a subset of bands or a single band is still arguable. We performed comprehensive analysis using individual bands and different subsets of bands to determine the effectiveness of spectral information for determining the anomaly in colorectal images. Our multispectral colorectal dataset consists of four classes, each represented by infra-red spectrum bands in addition to the visual spectrum bands. We performed our analysis of spectral imaging by stratifying the abnormalities using both spatial and spectral information. For our experiments, we used a combination of texture descriptors with an ensemble classification approach that performed best on our dataset. We applied our method to another dataset and got comparable results with those obtained using the state-of-the-art method and convolutional neural network based method. Moreover, we explored the relationship of the number of bands with the problem complexity and found that higher number of bands is required for a complex task to achieve improved performance. Our results demonstrate a synergy between infra-red and visual spectrum by improving the classification accuracy (by 6%) on incorporating the infra-red representation. We also highlight the importance of how the dataset should be divided into training and testing set for evaluating the histopathology image-based approaches, which has not been considered in previous studies on multispectral histopathology images.

The following information is missing from the Funding section: This article was funded by the Qatar National Library. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.