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Native fluorescence spectra are acquired from fresh normal and cancerous human prostate tissues. The fluorescence data are analyzed using an unsupervised machine learning algorithm such as non-negative matrix factorization. The nonnegative spectral components are retrieved and attributed to the native fluorophores such as collagen, reduced nicotinamide adenine dinucleotide (NADH), and flavin adenine dinucleotide (FAD) in tissue. The retrieved scores of the components are used to estimate the relative concentrations of the native fluorophores such as NADH and FAD and the redox ratio. A supervised machine learning algorithm such as support vector machine (SVM) is used to classify normal and cancerous tissue samples based on either the relative concentrations of NADH and FAD or the redox ratio alone. Various statistical measures such as sensitivity, specificity, and accuracy, along with the area under receiver operating characteristic (ROC) curve are used to show the classification performance. A cross validation method such as leave-one-out is used to further evaluate the predictive performance of the SVM classifier to avoid bias due to overfitting, and the accuracy was found to be 93.3%.

Food spoilage is mainly caused by microorganisms, such as bacteria. In this study, we measure the autofluorescence in meat samples longitudinally over a week in an attempt to develop a method to rapidly detect meat spoilage using fluorescence spectroscopy. Meat food is a biological tissue, which contains intrinsic fluorophores, such as tryptophan, collagen, nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) etc. As meat spoils, it undergoes various morphological and chemical changes. The concentrations of the native fluorophores present in a sample may change. In particular, the changes in NADH and FAD are associated with microbial metabolism, which is the most important process of the bacteria in food spoilage. Such changes may be revealed by fluorescence spectroscopy and used to indicate the status of meat spoilage. Therefore, such native fluorophores may be unique, reliable and non-subjective indicators for detection of spoiled meat. The results of the study show that the relative concentrations of all above fluorophores change as the meat samples kept in room temperature (~19°C) spoil. The changes become more rapidly after about two days. For the meat samples kept in a freezer (~ -12°C), the changes are much less or even unnoticeable over a-week-long storage.

Context.-Distinguishing chromophobe renal cell carcinoma (chRCC), especially in the presence of eosinophilic cytoplasm, from oncocytoma on hematoxylin-eosin can be difficult and often requires time-consuming ancillary procedures that ultimately may not be informative. Objective.-To explore the potential of multiphoton microscopy (MPM) as an alternative and rapid diagnostic tool in differentiating oncocytoma from chRCC at subcellular resolution without tissue processing. Design.-Unstained, deparaffinized tissue sections from 27 tumors (oncocytoma [n = 12], chRCC [n = 12], eosinophilic variant of chRCC [n = 1], and atypical oncocytic renal neoplasm [n = 2]) were imaged with MPM. Morphologic evaluation and automated quantitative morphometric analysis were conducted to distinguish between chRCC and oncocytoma. Results.-The typical cases of oncocytomas (12 of 12) and chRCC (12 of 12) could be readily differentiated on MPM based on the morphologic features similar to hematoxylin-eosin. The most striking MPM signature of both of the tumors was the presence of autofluorescent intracytoplasmic granules, which are not seen on hematoxylin-eosin-stained slides. Although we saw these granules in both types of tumors, they appeared distinct, based on their size, shape, cytoplasmic distribution, and autofluorescence wavelengths, and were valuable in arriving at a definitive diagnosis. For oncocytomas and chRCC, high diagnostic accuracies of 100% and 83.3% were achieved on blinded MPM and morphometric analysis, respectively. Conclusions.-To the best of our knowledge, this is the first demonstration of MPM to distinguish chRCC from oncocytoma in fixed tissues. Our study was limited by small sample size and only a few variants of oncocytic tumors. Prospective studies are warranted to assess the utility of MPM as a diagnostic aid in oncocytic renal tumors. © Copyright 2018 College of American Pathologists.

Worldwide breast cancer incidence has increased by more than twenty percent in the past decade. It is also known that in that time, mortality due to the affliction has increased by fourteen percent. Using optical-based diagnostic techniques, such as Raman spectroscopy, has been explored in order to increase diagnostic accuracy in a more objective way along with significantly decreasing diagnostic wait-times. In this study, Raman spectroscopy with 532-nm excitation was used in order to incite resonance effects to enhance Stokes Raman scattering from unique biomolecular vibrational modes. Seventy-two Raman spectra (41 cancerous, 31 normal) were collected from nine breast tissue samples by performing a ten-spectra average using a 500-ms acquisition time at each acquisition location. The raw spectral data was subsequently prepared for analysis with background correction and normalization. The spectral data in the Raman Shift range of 750- 2000 cm-1 was used for analysis since the detector has highest sensitivity around in this range. The matrix decomposition technique nonnegative matrix factorization (NMF) was then performed on this processed data. The resulting leave-oneout cross-validation using two selective feature components resulted in sensitivity, specificity and accuracy of 92.6%, 100% and 96.0% respectively. The performance of NMF was also compared to that using principal component analysis (PCA), and NMF was shown be to be superior to PCA in this study. This study shows that coupling the resonance Raman spectroscopy technique with subsequent NMF decomposition method shows potential for high characterization accuracy in breast cancer detection.

A clear distinction between oncocytoma and chromophobe renal cell carcinoma (chRCC) is critically important for clinical management of patients. But it may often be difficult to distinguish the two entities based on hematoxylin and eosin (H and E) stained sections alone. In this study, second harmonic generation (SHG) signals which are very specific to collagen were used to image collagen fibril structure. We conduct a pilot study to develop a new diagnostic method based on the analysis of collagen associated with kidney tumors using convolutional neural networks (CNNs). CNNs comprise a type of machine learning process well-suited for drawing information out of images. This study examines a CNN model's ability to differentiate between oncocytoma (benign), and chRCC (malignant) kidney tumor images acquired with second harmonic generation (SHG), which is very specific for collagen matrix. To the best of our knowledge, this is the first study that attempts to distinguish the two entities based on their collagen structure. The model developed from this study demonstrated an overall classification accuracy of 68.7% with a specificity of 66.3% and sensitivity of 74.6%. While these results reflect an ability to classify the kidney tumors better than chance, further studies will be carried out to (a) better realize the tumor classification potential of this method with a larger sample size and (b) combining SHG with two-photon excited intrinsic fluorescence signal to achieve better classification. © COPYRIGHT SPIE. Downloading of the abstract is permitted for personal use only.

Resonance Raman spectroscopy using 532nm excitation was used to distinguish normal brain tissue from different grades of glioma tissues. Principal component analysis was used to analyze the spectral data and achieved high accuracy. © 2018 The Author(s).

VRR spectroscopy was used for BCC and normal skin tissues with 532nm excitation. The spectra showed significant changes in collagen, carotenoids and lipids. These enhanced fingerprints demonstrate a potential use as label-free pathology method. © 2018 The Author(s).

The goal of the research is to determine the prognostic molecular pathological changes in components and composition, for human brain glioma gradings in comparison with normal tissues in three-dimensional Raman imaging profiles by visible Resonance Raman (VRR) imaging. <p> </p>VRR images from twenty-five specimens including three healthy tissues, one normal control, and twenty-one glioma tissues of grades II, II-III and III-IV with histology examination were measured and investigated using WITec300R confocal micro Raman imaging system with laser excitation of 532nm. <p> </p>Two-dimensional RR spectral mappings performed in 20&mu;m x 20&mu;m generated 400 images which integrated the intensity of the specific biochemical bonds as the third dimension. The three-dimension (3D) map demonstrated the spatial distributions of three selected sets of RR spectra of molecular biomarkers, and revealed significant differences in the spectra between normal and glioma tissues of different grades due to the composition changes in key molimageecules. These RR molecular spectral fingerprints have displayed: a clear enhancement of RR vibrational modes at 1129-1131cm^-1 and 2934cm^-1 which are supposed to be arising from lipoproteins; evident decreased RR vibrational modes at 1442cm^-1 and 2854cm^-1 which are from saturated fatty acids bonds in all-grades of glioma brain tissues compared with normal tissues; and the enhanced RR spectral modes of 1129 cm^-1 and 2938cm^-1 which suggest contribution from lactate. These findings may provide a novel proof for anaerobic glycolysis metabolic process in brain glioma cancer tissues that has been explained by Warburg effects.

Analyzing spectral or imaging data collected with various optical biopsy methods is often times difficult due to the complexity of the biological basis. Robust methods that can utilize the spectral or imaging data and detect the characteristic spectral or spatial signatures for different types of tissue is challenging but highly desired. In this study, we used various machine learning algorithms to analyze a spectral dataset acquired from human skin normal and cancerous tissue samples using resonance Raman spectroscopy with 532nm excitation. The algorithms including principal component analysis, nonnegative matrix factorization, and autoencoder artificial neural network are used to reduce dimension of the dataset and detect features. A support vector machine with a linear kernel is used to classify the normal tissue and cancerous tissue samples. The efficacies of the methods are compared.

The accurate identification of the human brain tumor boundary and the complete resection of the tumor are two essential factors for the removal of the glioma tumor in brain surgery. We present a visible resonance Raman (VRR) spectroscopy technique for differentiating the brain tumor margin and glioma grading. Eighty-seven VRR spectra from twenty-one human brain specimens of four types of brain tissues, including the control, glioma grade II, III, and IV tissues, were observed. This study focuses on observing the characteristics of new biomarkers and their changes in the four types of brain tissue. We found that two new RR peaks at 1129 cm-1 and 1338 cm-1 associated with molecular vibrational bonds in four types of brain tissues are significantly different in peak intensities of VRR spectra. These two resonance enhanced peaks may arise from lactic acid/phosphatidic acid and adenosine triphosphate (ATP)/nicotinamide adenine dinucleotide, respectively. We found that lactic acid and ATP concentrations vary with glioma gratings. The higher the grade of malignancy, the more the increase in lactic acid and ATP concentrations. These two RR peaks may be considered as new molecular biomarkers and used to evaluate glioma grades and identify the margin of gliomas from the control tissues. The metabolic process of lactic acid and ATP in glioma cells based on the VRR spectral changes may reveal the Warburg hypothesis. © 2018 Author(s).

Visible resonant Raman (VRR) spectroscopy provides an effective way to enhance Raman signal from particular bonds associated with key molecules due to changes on molecular level. This paper reports on the VRR use for detection of human brain the control and gliomas of three grades. From the RR spectra additional two molecular vibrational biomarkers at 1129cm^-1 and 1338cm^-1, for the four types of brain tissues are significantly different in intensity. The new RR spectral peaks can be used as molecular biomarkers to evaluate glioma grades and identify the margin of gliomas from the controls. The metabolic process of glioma cells based on the RR spectral changes may reveal the Warburg hypothesis.