We present a rapid, portable optical system for label-free detection of COVID-19. Raman spectra from an entire liquid drop of saliva supernatant can be obtained within 6 minutes, and the sample is classified as COVID-19 positive or negative using artificial intelligence (AI).
293 COVID negative and 49 COVID positive saliva supernatant samples were analyzed. Positive samples were from hospitalized patients (non-critical and critical) and non-hospitalized testing clinic volunteers (symptomatic and asymptomatic). Our Raman/AI system has an 82% accuracy detecting people with COVID-19 of any severity with any symptom presentation, and 89% accuracy when detecting COVID-19 in hospitalized patients alone. Rapid label-free analysis of biofluids for viruses could provide a low-cost screening solution that could be adapted to respond to viral mutations. This could be invaluable for future pandemics and for reducing infections in hospitals, care homes and workplaces.
SignificanceAs many as 60% of patients with early stage breast cancer undergo breast-conserving surgery. Of those, 20% to 35% need a second surgery because of incomplete resection of the lesions. A technology allowing in situ detection of cancer could reduce re-excision procedure rates and improve patient survival.AimRaman spectroscopy was used to measure the spectral fingerprint of normal breast and cancer tissue ex-vivo. The aim was to build a machine learning model and to identify the biomolecular bands that allow one to detect invasive breast cancer.ApproachThe system was used to interrogate specimens from 20 patients undergoing lumpectomy, mastectomy, or breast reduction surgery. This resulted in 238 ex-vivo measurements spatially registered with standard histology classifying tissue as cancer, normal, or fat. A technique based on support vector machines led to the development of predictive models, and their performance was quantified using a receiver-operating-characteristic analysis.ResultsRaman spectroscopy combined with machine learning detected normal breast from ductal or lobular invasive cancer with a sensitivity of 93% and a specificity of 95%. This was achieved using a model based on only two spectral bands, including the peaks associated with C–C stretching of proteins around 940 cm − 1 and the symmetric ring breathing at 1004 cm − 1 associated with phenylalanine.ConclusionsDetection of cancer on the margins of surgically resected breast specimen is feasible with Raman spectroscopy.
SignificanceStandardized data processing approaches are required in the field of bio-Raman spectroscopy to ensure information associated with spectral data acquired by different research groups, and with different systems, can be compared on an equal footing.AimAn open-sourced data processing software package was developed, implementing algorithms associated with all steps required to isolate the inelastic scattering component from signals acquired using Raman spectroscopy devices. The package includes a novel morphological baseline removal technique (BubbleFill) that provides increased adaptability to complex baseline shapes compared to current gold standard techniques. Also incorporated in the package is a versatile tool simulating spectroscopic data with varying levels of Raman signal-to-background ratios, baselines with different morphologies, and varying levels of stochastic noise.ResultsApplication of the BubbleFill technique to simulated data demonstrated superior baseline removal performance compared to standard algorithms, including iModPoly and MorphBR. The data processing workflow of the open-sourced package was validated in four independent in-human datasets, demonstrating it leads to inter-systems data compatibility.ConclusionsA new open-sourced spectroscopic data pre-processing package was validated on simulated and real-world in-human data and is now available to researchers and clinicians for the development of new clinical applications using Raman spectroscopy.
Significance: The diagnosis and treatment of prostate cancer (PCa) are limited by a lack of intraoperative information to accurately target tumors with needles for biopsy and brachytherapy. An innovative image-guidance technique using optical devices could improve the diagnostic yield of biopsy and efficacy of radiotherapy.Aim: To evaluate the performance of multimodal PCa detection using biomolecular features from in-situ Raman spectroscopy (RS) combined with image-based (radiomics) features from multiparametric magnetic resonance images (mpMRI).Approach: In a prospective pilot clinical study, 18 patients were recruited and underwent high-dose-rate brachytherapy. Multimodality image fusion (preoperative mpMRI with intraoperative transrectal ultrasound) combined with electromagnetic tracking was used to navigate an RS needle in the prostate prior to brachytherapy. This resulting dataset consisted of Raman spectra and co-located radiomics features from mpMRI. Feature selection was performed with the constraint that no more than 10 features were retained overall from a combination of inelastic scattering spectra and radiomics. These features were used to train support vector machine classifiers for PCa detection based on leave-one-patient-out cross-validation.Results: RS along with biopsy samples were acquired from 47 sites along the insertion trajectory of the fiber-optics needle: 26 were confirmed as benign or grade group = 1, and 21 as grade group >1, according to histopathological reports. The combination of the fingerprint region of the RS and radiomics showed an accuracy of 83% (sensitivity = 81 % and a specificity = 85 % ), outperforming by more than 9% models trained with either spectroscopic or mpMRI data alone. An optimal number of features was identified between 6 and 8 features, which have good potential for discriminating grade group ≥1 / grade group <1 (accuracy = 87 % ) or grade group >1 / grade group ≤1 (accuracy = 91 % ).Conclusions: In-situ Raman spectroscopy combined with mpMRI radiomics features can lead to highly accurate PCa detection for improved in-vivo targeting of biopsy sample collection and radiotherapy seed placement.
Significance: The diagnosis of prostate cancer (PCa) and focal treatment by brachytherapy are limited by the lack of precise intraoperative information to target tumors during biopsy collection and radiation seed placement. Image-guidance techniques could improve the safety and diagnostic yield of biopsy collection as well as increase the efficacy of radiotherapy.Aim: To estimate the accuracy of PCa detection using in situ Raman spectroscopy (RS) in a pilot in-human clinical study and assess biochemical differences between in vivo and ex vivo measurements.Approach: A new miniature RS fiber-optics system equipped with an electromagnetic (EM) tracker was guided by trans-rectal ultrasound-guided imaging, fused with preoperative magnetic resonance imaging to acquire 49 spectra in situ (in vivo) from 18 PCa patients. In addition, 179 spectra were acquired ex vivo in fresh prostate samples from 14 patients who underwent radical prostatectomy. Two machine-learning models were trained to discriminate cancer from normal prostate tissue from both in situ and ex vivo datasets.Results: A support vector machine (SVM) model was trained on the in situ dataset and its performance was evaluated using leave-one-patient-out cross validation from 28 normal prostate measurements and 21 in-tumor measurements. The model performed at 86% sensitivity and 72% specificity. Similarly, an SVM model was trained with the ex vivo dataset from 152 normal prostate measurements and 27 tumor measurements showing reduced cancer detection performance mostly attributable to spatial registration inaccuracies between probe measurements and histology assessment. A qualitative comparison between in situ and ex vivo measurements demonstrated a one-to-one correspondence and similar ratios between the main Raman bands (e.g., amide I-II bands, phenylalanine).Conclusions: PCa detection can be achieved using RS and machine learning models for image-guidance applications using in situ measurements during prostate biopsy procedures.
Significance: The primary method of COVID-19 detection is reverse transcription polymerase chain reaction (RT-PCR) testing. PCR test sensitivity may decrease as more variants of concern arise and reagents may become less specific to the virus.
Aim: We aimed to develop a reagent-free way to detect COVID-19 in a real-world setting with minimal constraints on sample acquisition. The machine learning (ML) models involved could be frequently updated to include spectral information about variants without needing to develop new reagents.
Approach: We present a workflow for collecting, preparing, and imaging dried saliva supernatant droplets using a non-invasive, label-free technique—Raman spectroscopy—to detect changes in the molecular profile of saliva associated with COVID-19 infection.
Results: We used an innovative multiple instance learning-based ML approach and droplet segmentation to analyze droplets. Amongst all confounding factors, we discriminated between COVID-positive and COVID-negative individuals yielding receiver operating coefficient curves with an area under curve (AUC) of 0.8 in both males (79% sensitivity and 75% specificity) and females (84% sensitivity and 64% specificity). Taking the sex of the saliva donor into account increased the AUC by 5%.
Conclusion: These findings may pave the way for new rapid Raman spectroscopic screening tools for COVID-19 and other infectious diseases.
Significance: Prostate cancer is the most common cancer among men. An accurate diagnosis of its severity at detection plays a major role in improving their survival. Recently, machine learning models using biomarkers identified from Raman micro-spectroscopy discriminated intraductal carcinoma of the prostate (IDC-P) from cancer tissue with a ≥85 % detection accuracy and differentiated high-grade prostatic intraepithelial neoplasia (HGPIN) from IDC-P with a ≥97.8 % accuracy.
Aim: To improve the classification performance of machine learning models identifying different types of prostate cancer tissue using a new dimensional reduction technique.
Approach: A radial basis function (RBF) kernel support vector machine (SVM) model was trained on Raman spectra of prostate tissue from a 272-patient cohort (Centre hospitalier de l’Université de Montréal, CHUM) and tested on two independent cohorts of 76 patients [University Health Network (UHN)] and 135 patients (Centre hospitalier universitaire de Québec-Université Laval, CHUQc-UL). Two types of engineered features were used. Individual intensity features, i.e., Raman signal intensity measured at particular wavelengths and novel Raman spectra fitted peak features consisting of peak heights and widths.
Results: Combining engineered features improved classification performance for the three aforementioned classification tasks. The improvements for IDC-P/cancer classification for the UHN and CHUQc-UL testing sets in accuracy, sensitivity, specificity, and area under the curve (AUC) are (numbers in parenthesis are associated with the CHUQc-UL testing set): +4 % (+8 % ), +7 % (+9 % ), +2 % (6%), +9 (+9) with respect to the current best models. Discrimination between HGPIN and IDC-P was also improved in both testing cohorts: +2.2 % (+1.7 % ), +4.5 % (+3.6 % ), +0 % (+0 % ), +2.3 (+0). While no global improvements were obtained for the normal versus cancer classification task [+0 % (−2 % ), +0 % (−3 % ), +2 % (−2 % ), +4 (+3)], the AUC was improved in both testing sets.
Conclusions: Combining individual intensity features and novel Raman fitted peak features, improved the classification performance on two independent and multicenter testing sets in comparison to using only individual intensity features.
KEYWORDS: Raman spectroscopy, Tissues, Signal to noise ratio, In vivo imaging, Cancer, Brain, Data acquisition, Luminescence, Tissue optics, Visualization
Significance: Ensuring spectral quality is prerequisite to Raman spectroscopy applied to surgery. This is because the inclusion of poor-quality spectra in the training phase of Raman-based pathology detection models can compromise prediction robustness and generalizability to new data. Currently, there exists no quantitative spectral quality assessment technique that can be used to either reject low-quality data points in existing Raman datasets based on spectral morphology or, perhaps more importantly, to optimize the in vivo data acquisition process to ensure minimal spectral quality standards are met.
Aim: To develop a quantitative method evaluating Raman signal quality based on the variance associated with stochastic noise in important tissue bands, including C─C stretch, CH2 / CH3 deformation, and the amide bands.
Approach: A single-point hand-held Raman spectroscopy probe system was used to acquire 315 spectra from 44 brain cancer patients. All measurements were classified as either high or low quality based on visual assessment (qualitative) and using a quantitative quality factor (QF) metric. Receiver-operator-characteristic (ROC) analyses were performed to evaluate the performance of the quantitative metric to assess spectral quality and improve cancer detection accuracy.
Results: The method can separate high- and low-quality spectra with a sensitivity of 89% and a specificity of 90% which is shown to increase cancer detection sensitivity and specificity by up to 20% and 12%, respectively.
Conclusions: The QF threshold is effective in stratifying spectra in terms of spectral quality and the observed false negatives and false positives can be linked to limitations of qualitative spectral quality assessment.
Epilepsy is a neurological disorder characterized by chronic excessive neuronal discharges. Epilepsy surgery may be considered in patients who are resistant to drug treatment, and several tests are used during pre-operative planning to locate the epileptic focus to be resected (MRI, PET, SPECT, EEG, MEG). In some cases, intracranial EEG monitoring or intraoperative electrocorticography is required to confirm and better delineate the area to be resected. Despite available tests, epilepsy surgery outcome remains modest. We present a multimodal imaging platform connected to a neurosurgical microscope allowing intraoperative detection of intrinsic optical brain biomarkers for surgical guidance during epilepsy surgery. Hyperspectral imaging (HSI) allows detection of the hemodynamic response associated with epileptic activity, spatial frequency domain imaging (SFDI) is used for optical properties reconstruction (absorption and reduced scattering), and auto-fluorescence imaging (AFI) allows metabolic markers identification. Validation of SFDI and AFI systems was performed on optical phantoms. Acquisitions on ex vivo tissue samples demonstrate the capabilities of the system to produce fluorescence intensity maps, calibrated with optical properties obtained from SFDI to account for tissue attenuation. In vivo acquisition during epilepsy surgery with HSI allows characterization of the hemodynamic response associated with epileptic spikes.
Prostate cancer is the most diagnosed form of cancer among American men and, in vast proportion, the standard of care treatment includes radical prostatectomy. Important risk factors associated with prostatectomies are the presence of post-surgery residual prostate tissue and positive cancer margins, potentially leading to recurrences. Prostate histopathology analysis following the procedure is used to determine follow-up treatment. However, only a limited fraction of the prostate margins can be sampled, which can lead to suboptimal evaluation and treatment. Here we present the development of a wide-field multimodal imaging system designed to quantify intrinsic tissue fluorescence and map scattering and absorption coefficients using spatial frequency domain imaging (SFDI). The system allows targeting of suspicious prostate regions to guide histopathology analysis, aiming to improve diagnostic accuracy and treatment planning. Tissue excitation for endogenous fluorescence is achieved with a 405 nm laser diode and, for SFDI, a digital light projector transmits structured white light used to reconstruct tissue optical properties (absorption, scattering) between 420 and 720 nm. A light transport model-based quantification algorithm then corrects the fluorescence spectra for tissue attenuation, lending a biomarker that correlates with local fluorophore concentrations. Spectral and spatial calibration of both modalities was done on optical phantoms and validation of the fluorescence quantification on biological tissue. Finally, imaging results are presented for 5 human prostates interrogated with the system, along with spatially-registered histopathology analyses. Future work involves massive data acquisition and development of artificial intelligence models for tissue classification (prostate, non-prostate; healthy, cancerous) and adaptation for intraoperative use.
Surgical excision of the whole prostate through a radical prostatectomy procedure is part of the standard of care for prostate cancer. Positive surgical margins (cancer cells having spread into surrounding nonresected tissue) occur in as many as 1 in 5 cases and strongly correlate with disease recurrence and the requirement of adjuvant treatment. Margin assessment is currently only performed by pathologists hours to days following surgery and the integration of a real-time surgical readout would benefit current prostatectomy procedures. Raman spectroscopy is a promising technology to assess surgical margins: its in vivo use during radical prostatectomy could help insure the extent of resected prostate and cancerous tissue is maximized. We thus present the design and development of a dual excitation Raman spectroscopy system (680- and 785-nm excitations) integrated to the robotic da Vinci surgical platform for in vivo use. Following validation in phantoms, spectroscopic data from 20 whole human prostates immediately following radical prostatectomy are obtained using the system. With this dataset, we are able to distinguish prostate from extra prostatic tissue with an accuracy, sensitivity, and specificity of 91%, 90.5%, and 96%, respectively. Finally, the integrated Raman spectroscopy system is used to collect preliminary spectroscopic data at the surgical margin in vivo in four patients.
Optical coherence tomography (OCT) yields microscopic volumetric images representing tissue structures based on the contrast provided by elastic light scattering. Multipatient studies using OCT for detection of tissue abnormalities can lead to large datasets making quantitative and unbiased assessment of classification algorithms performance difficult without the availability of automated analytical schemes. We present a mathematical descriptor reducing the dimensionality of a classifier’s input data, while preserving essential volumetric features from reconstructed three-dimensional optical volumes. This descriptor is used as the input of classification algorithms allowing a detailed exploration of the features space leading to optimal and reliable classification models based on support vector machine techniques. Using imaging dataset of paraffin-embedded tissue samples from 38 ovarian cancer patients, we report accuracies for cancer detection >90% for binary classification between healthy fallopian tube and ovarian samples containing cancer cells. Furthermore, multiples classes of statistical models are presented demonstrating >70% accuracy for the detection of high-grade serous, endometroid, and clear cells cancers. The classification approach reduces the computational complexity and needed resources to achieve highly accurate classification, making it possible to contemplate other applications, including intraoperative surgical guidance, as well as other depth sectioning techniques for fresh tissue imaging.
Prostate cancer is the most frequent diagnosed cancers among men. When prostate cancer occurs, the cancer does not result in only one or few localized malignant tumor, but is generally spread within the whole prostate.
In order to counteract the very high level of heterogeneities exhibited by prostate tissues, we developed a method for high-resolution co-registration of Raman spectroscopy with prostate cancer diagnosis.
Raman spectra were acquired on fresh ex vivo prostate within 2 hours after radical prostatectomy using a multi-wavelength hand-held contact probe. After the measurements, the prostate was reintegrated to the usual pathological workflow: formalin fixated and paraffin embedded (FFPE), and prepared for microscope histopathological analyses. The precise reconstruction of the prostate slice with hematoxylin and eosin (H and E) tissue allows the spatial correlation of the measured area (0.2 mm2) with the correspondent histopathological information, for point-by-point diagnosis determination. The tissue was classified into groups (normal/cancer) and subgroups according to the percentage of benign glands, stroma or cancer.
Different machine learning algorithms were tested to classify the spectra with increasing levels of categorization. Preliminary results showed that Raman spectroscopy is capable of detecting prostate cancer with an accuracy >90%. In addition, high percentages of stroma (vs. glands) have been correlated with spectral signature of collagen, which is the main constituent of extracellular matrix.
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