Broadband hyperspectral imaging for breast tumor detection using spectral and spatial information

Complete tumor removal during breast-conserving surgery remains challenging due to the lack of optimal intraoperative margin assessment techniques. Here, we use hyperspectral imaging for tumor detection in fresh breast tissue. We evaluated different wavelength ranges and two classification algorithms; a pixel-wise classification algorithm and a convolutional neural network that combines spectral and spatial information. The highest classification performance was obtained using the full wavelength range (450-1650nm). Adding spatial information mainly improved the differentiation of tissue classes within the malignant and healthy classes. High sensitivity and specificity were accomplished, which offers potential for hyperspectral imaging as a margin assessment technique to improve surgical outcome. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreemen

Method for coregistration of optical measurements of breast tissue with histopathology : the importance of accounting for tissue deformations

For the validation of optical diagnostic technologies, experimental results need to be benchmarked against the gold standard. Currently, the gold standard for tissue characterization is assessment of hematoxylin and eosin (H&E)-stained sections by a pathologist. When processing tissue into H&E sections, the shape of the tissue deforms with respect to the initial shape when it was optically measured. We demonstrate the importance of accounting for these tissue deformations when correlating optical measurement with routinely acquired histopathology. We propose a method to register the tissue in the H&E sections to the optical measurements, which corrects for these tissue deformations. We compare the registered H&E sections to H&E sections that were registered with an algorithm that does not account for tissue deformations by evaluating both the shape and the composition of the tissue and using microcomputer tomography data as an independent measure. The proposed method, which did account for tissue deformations, was more accurate than the method that did not account for tissue deformations. These results emphasize the need for a registration method that accounts for tissue deformations, such as the method presented in this study, which can aid in validating optical techniques for clinical use. (C) The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License

A comparison of spectroscopy and imaging techniques utilizing spectrally resolved diffusely reflected light for intraoperative margin assessment in breast-conserving surgery: a systematic review and meta-analysis

Up to 19% of patients require re-excision surgery due to positive margins in breast-conserving surgery (BCS). Intraoperative margin assessment tools (IMAs) that incorporate tissue optical measurements could help reduce re-excision rates. This review focuses on methods that use and assess spectrally resolved diffusely reflected light for breast cancer detection in the intraoperative setting. Following PROSPERO registration (CRD42022356216), an electronic search was performed. The modalities searched for were diffuse reflectance spectroscopy (DRS), multispectral imaging (MSI), hyperspectral imaging (HSI), and spatial frequency domain imaging (SFDI). The inclusion criteria encompassed studies of human in vivo or ex vivo breast tissues, which presented data on accuracy. The exclusion criteria were contrast use, frozen samples, and other imaging adjuncts. 19 studies were selected following PRISMA guidelines. Studies were divided into point-based (spectroscopy) or whole field-of-view (imaging) techniques. A fixed-or random-effects model analysis generated pooled sensitivity/specificity for the different modalities, following heterogeneity calculations using the Q statistic. Overall, imaging-based techniques had better pooled sensitivity/specificity (0.90 (CI 0.76-1.03)/0.92 (CI 0.78-1.06)) compared with probe-based techniques (0.84 (CI 0.78-0.89)/0.85 (CI 0.79-0.91)). The use of spectrally resolved diffusely reflected light is a rapid, non-contact technique that confers accuracy in discriminating between normal and malignant breast tissue, and it constitutes a potential IMA tool

Framework for Hyperspectral Image Processing and Quantification for Cancer Detection During Animal Tumor Surgery

Hyperspectral imaging (HSI) is an imaging modality that holds strong potential for rapid cancer detection during image-guided surgery. But the data from HSI often needs to be processed appropriately in order to extract the maximum useful information that differentiates cancer from normal tissue. We proposed a framework for hyperspectral image processing and quantification, which includes a set of steps including image preprocessing, glare removal, feature extraction, and ultimately image classification. The framework has been tested on images from mice with head and neck cancer, using spectra from 450- to 900-nm wavelength. The image analysis computed Fourier coefficients, normalized reflectance, mean, and spectral derivatives for improved accuracy. The experimental results demonstrated the feasibility of the hyperspectral image processing and quantification framework for cancer detection during animal tumor surgery, in a challenging setting where sensitivity can be low due to a modest number of features present, but potential for fast image classification can be high. This HSI approach may have potential application in tumor margin assessment during image-guided surgery, where speed of assessment may be the dominant factor

Intra-operative spectroscopic assessment of surgical margins during breast conserving surgery

Background: In over 20% of breast conserving operations, postoperative pathological assessment of the excised tissue reveals positive margins, requiring additional surgery. Current techniques for intra-operative assessment of tumor margins are insufficient in accuracy or resolution to reliably detect small tumors. There is a distinct need for a fast technique to accurately identify tumors smaller than 1 mm2 in large tissue surfaces within 30 min. Methods: Multi-modal spectral histopathology (MSH), a multimodal imaging technique combining tissue auto-fluorescence and Raman spectroscopy was used to detect microscopic residual tumor at the surface of the excised breast tissue. New algorithms were developed to optimally utilize auto-fluorescence images to guide Raman measurements and achieve the required detection accuracy over large tissue surfaces (up to 4 × 6.5 cm2). Algorithms were trained on 91 breast tissue samples from 65 patients. Results: Independent tests on 121 samples from 107 patients - including 51 fresh, whole excision specimens - detected breast carcinoma on the tissue surface with 95% sensitivity and 82% specificity. One surface of each uncut excision specimen was measured in 12–24 min. The combination of high spatial-resolution auto-fluorescence with specific diagnosis by Raman spectroscopy allows reliable detection even for invasive carcinoma or ductal carcinoma in situ smaller than 1 mm2. Conclusions: This study provides evidence that this multimodal approach could provide an objective tool for intra-operative assessment of breast conserving surgery margins, reducing the risk for unnecessary second operations

Development of deep learning methods for head and neck cancer detection in hyperspectral imaging and digital pathology for surgical guidance

Surgeons performing routine cancer resections utilize palpation and visual inspection, along with time-consuming microscopic tissue analysis, to ensure removal of cancer. Despite this, inadequate surgical cancer margins are reported for up to 10-20% of head and neck squamous cell carcinoma (SCC) operations. There exists a need for surgical guidance with optical imaging to ensure complete cancer resection in the operating room. The objective of this dissertation is to evaluate hyperspectral imaging (HSI) as a non-contact, label-free optical imaging modality to provide intraoperative diagnostic information. For comparison of different optical methods, autofluorescence, RGB composite images synthesized from HSI, and two fluorescent dyes are also acquired and investigated for head and neck cancer detection. A novel and comprehensive dataset was obtained of 585 excised tissue specimens from 204 patients undergoing routine head and neck cancer surgeries. The first aim was to use SCC tissue specimens to determine the potential of HSI for surgical guidance in the challenging task of head and neck SCC detection. It is hypothesized that HSI could reduce time and provide quantitative cancer predictions. State-of-the-art deep learning algorithms were developed for SCC detection in 102 patients and compared to other optical methods. HSI detected SCC with a median AUC score of 85%, and several anatomical locations demonstrated good SCC detection, such as the larynx, oropharynx, hypopharynx, and nasal cavity. To understand the ability of HSI for SCC detection, the most important spectral features were calculated and correlated with known cancer physiology signals, notably oxygenated and deoxygenated hemoglobin. The second aim was to evaluate HSI for tumor detection in thyroid and salivary glands, and RGB images were synthesized using the spectral response curves of the human eye for comparison. Using deep learning, HSI detected thyroid tumors with 86% average AUC score, which outperformed fluorescent dyes and autofluorescence, but HSI-synthesized RGB imagery performed with 90% AUC score. The last aim was to develop deep learning algorithms for head and neck cancer detection in hundreds of digitized histology slides. Slides containing SCC or thyroid carcinoma can be distinguished from normal slides with 94% and 99% AUC scores, respectively, and SCC and thyroid carcinoma can be localized within whole-slide images with 92% and 95% AUC scores, respectively. In conclusion, the outcomes of this thesis work demonstrate that HSI and deep learning methods could aid surgeons and pathologists in detecting head and neck cancers.Ph.D

Histopathological image analysis : a review

Over the past decade, dramatic increases in computational power and improvement in image analysis algorithms have allowed the development of powerful computer-assisted analytical approaches to radiological data. With the recent advent of whole slide digital scanners, tissue histopathology slides can now be digitized and stored in digital image form. Consequently, digitized tissue histopathology has now become amenable to the application of computerized image analysis and machine learning techniques. Analogous to the role of computer-assisted diagnosis (CAD) algorithms in medical imaging to complement the opinion of a radiologist, CAD algorithms have begun to be developed for disease detection, diagnosis, and prognosis prediction to complement the opinion of the pathologist. In this paper, we review the recent state of the art CAD technology for digitized histopathology. This paper also briefly describes the development and application of novel image analysis technology for a few specific histopathology related problems being pursued in the United States and Europe

Advancing fluorescent contrast agent recovery methods for surgical guidance applications

Fluorescence-guided surgery (FGS) utilizes fluorescent contrast agents and specialized optical instruments to assist surgeons in intraoperatively identifying tissue-specific characteristics, such as perfusion, malignancy, and molecular function. In doing so, FGS represents a powerful surgical navigation tool for solving clinical challenges not easily addressed by other conventional imaging methods. With growing translational efforts, major hurdles within the FGS field include: insufficient tools for understanding contrast agent uptake behaviors, the inability to image tissue beyond a couple millimeters, and lastly, performance limitations of currently-approved contrast agents in accurately and rapidly labeling disease. The developments presented within this thesis aim to address such shortcomings. Current preclinical fluorescence imaging tools often sacrifice either 3D scale or spatial resolution. To address this gap in high-resolution, whole-body preclinical imaging tools available, the crux of this work lays on the development of a hyperspectral cryo-imaging system and image-processing techniques to accurately recapitulate high-resolution, 3D biodistributions in whole-animal experiments. Specifically, the goal is to correct each cryo-imaging dataset such that it becomes a useful reporter for whole-body biodistributions in relevant disease models. To investigate potential benefits of seeing deeper during FGS, we investigated short-wave infrared imaging (SWIR) for recovering fluorescence beyond the conventional top few millimeters. Through phantom, preclinical, and clinical SWIR imaging, we were able to 1) validate the capability of SWIR imaging with conventional NIR-I fluorophores, 2) demonstrate the translational benefits of SWIR-ICG angiography in a large animal model, and 3) detect micro-dose levels of an EGFR-targeted NIR-I probe during a Phase 0 clinical trial. Lastly, we evaluated contrast agent performances for FGS glioma resection and breast cancer margin assessment. To evaluate glioma-labeling performance of untargeted contrast agents, 3D agent biodistributions were compared voxel-by-voxel to gold-standard Gd-MRI and pathology slides. Finally, building on expertise in dual-probe ratiometric imaging at Dartmouth, a 10-pt clinical pilot study was carried out to assess the technique’s efficacy for rapid margin assessment. In summary, this thesis serves to advance FGS by introducing novel fluorescence imaging devices, techniques, and agents which overcome challenges in understanding whole-body agent biodistributions, recovering agent distributions at greater depths, and verifying agents’ performance for specific FGS applications

Biophysical basis of skin cancer detection using Raman spectroscopy

The goal of this dissertation is to study the potential of Raman spectroscopy in improving the clinical diagnosis of skin cancer, including two main applications: noninvasive screening of melanoma skin cancer and surgical margin detection of nonmelanoma skin cancer. Skin cancer is the most common type of malignancy, accounting for over 5.4 million cases and 10 thousand deaths per year in the United States alone. Like most cancers, the current “gold standard” diagnosis relies on biopsy and histopathology, which is invasive, time-consuming, and costly. Moreover, large numbers of benign lesions are biopsied for melanoma diagnosis, resulting in substantial financial burden and patient discomfort. Therefore, an urgent need exists to develop a noninvasive, fast, and accurate method for skin cancer detection. The first part of the dissertation focuses on exploring the biophysical origin of in vivo melanoma detection. Our group has previously reported on the development of a clinical Raman spectroscopy system towards spectral biopsy of skin; however, the biochemical changes that Raman spectroscopy relies on for accurate melanoma diagnosis remained unclear. As a result, we proposed a biophysical inverse model to address this issue. To build the model, we established a custom confocal Raman microscope to extract in situ human skin constituents spanning normal and various diseased states. Our results indicate collagen, elastin, keratin, cell nucleus, triolein, ceramide, melanin, and water are the most important model components. Furthermore, collagen and triolein are the most relevant markers to discriminate malignant melanoma from benign nevi. The second part of the dissertation discusses the biophysical basis of nonmelanoma skin cancer margin delineation. We discovered the diagnostic markers to accurately differentiate tumor from normal skin, which is critical to maximize positive patient outcomes in skin cancer surgery. The biochemical changes derived from our model were highly correlated with histopathological diagnosis. We further demonstrated the feasibility of a superpixel acquisition approach for rapid classification of tumor boundaries in skin biopsies. Our results suggest Raman spectroscopy will be a powerful tool for intraoperative surgical guidanceBiomedical Engineerin