Simultaneous Dual-Wavelength Source Raman Spectroscopy with a Handheld Confocal Probe for Analysis of the Chemical Composition of In Vivo Human Skin.

Confocal Raman spectroscopy (CRS) is a powerful tool that has been widely used for biological tissue analysis because of its noninvasive nature, high specificity, and rich biochemical information. However, current commercial CRS systems suffer from limited detection regions (450-1750 cm-1), bulky sizes, nonflexibilities, slow acquisitions by consecutive excitations, and high costs if using a Fourier transform (FT) Raman spectroscopy with an InGaAs detector, which impede their adoption in clinics. In this study, we developed a portable CRS system with a simultaneous dual-wavelength source and a miniaturized handheld probe (120 mm × 60 mm × 50 mm) that can acquire spectra in both fingerprint (FP, 450-1750 cm-1) and high wavenumber (HW, 2800-3800 cm-1) regions simultaneously. An innovative design combining 671 and 785 nm lasers for simultaneous excitation through a compact and high-efficiency (>90%) wavelength combiner was implemented. Moreover, to decouple the fused FP and HW spectra, a first-of-its-kind precise Raman spectra separation algorithm (PRSSA) was developed based on the maximum a posteriori probability (MAP) estimate. The accuracy of spectra separation was greater than 99%, demonstrated in both phantom experiments and in vivo human skin measurements. The total data acquisition time was reduced by greater than 50% compared to other CRS systems. The results proved our proposed CRS system and PRSSA's superior capability in fast and ultrawideband spectra acquisition will significantly improve the integration of CRS in the clinical workflow.

A triple-network organization for the mouse brain.

The triple-network model of psychopathology is a framework to explain the functional and structural neuroimaging phenotypes of psychiatric and neurological disorders. It describes the interactions within and between three distributed networks: the salience, default-mode, and central executive networks. These have been associated with brain disorder traits in patients. Homologous networks have been proposed in animal models, but their integration into a triple-network organization has not yet been determined. Using resting-state datasets, we demonstrate conserved spatio-temporal properties between triple-network elements in human, macaque, and mouse. The model predictions were also shown to apply in a mouse model for depression. To validate spatial homologies, we developed a data-driven approach to convert mouse brain maps into human standard coordinates. Finally, using high-resolution viral tracers in the mouse, we refined an anatomical model for these networks and validated this using optogenetics in mice and tractography in humans. Unexpectedly, we find serotonin involvement within the salience rather than the default-mode network. Our results support the existence of a triple-network system in the mouse that shares properties with that of humans along several dimensions, including a disease condition. Finally, we demonstrate a method to humanize mouse brain networks that opens doors to fully data-driven trans-species comparisons.

An Investigation of Signal Preprocessing for Photoacoustic Tomography.

Photoacoustic tomography (PAT) is increasingly being used for high-resolution biological imaging at depth. Signal-to-noise ratios and resolution are the main factors that determine image quality. Various reconstruction algorithms have been proposed and applied to reduce noise and enhance resolution, but the efficacy of signal preprocessing methods which also affect image quality, are seldom discussed. We, therefore, compared common preprocessing techniques, namely bandpass filters, wavelet denoising, empirical mode decomposition, and singular value decomposition. Each was compared with and without accounting for sensor directivity. The denoising performance was evaluated with the contrast-to-noise ratio (CNR), and the resolution was calculated as the full width at half maximum (FWHM) in both the lateral and axial directions. In the phantom experiment, counting in directivity was found to significantly reduce noise, outperforming other methods. Irrespective of directivity, the best performing methods for denoising were bandpass, unfiltered, SVD, wavelet, and EMD, in that order. Only bandpass filtering consistently yielded improvements. Significant improvements in the lateral resolution were observed using directivity in two out of three acquisitions. This study investigated the advantages and disadvantages of different preprocessing methods and may help to determine better practices in PAT reconstruction.

Machine Learning Assisted Handheld Confocal Raman Micro-Spectroscopy for Identification of Clinically Relevant Atopic Eczema Biomarkers.

Atopic dermatitis (AD) is a common chronic inflammatory skin dermatosis condition due to skin barrier dysfunction that causes itchy, red, swollen, and cracked skin. Currently, AD severity clinical scores are subjected to intra- and inter-observer differences. There is a need for an objective scoring method that is sensitive to skin barrier differences. The aim of this study was to evaluate the relevant skin chemical biomarkers in AD patients. We used confocal Raman micro-spectroscopy and advanced machine learning methods as means to classify eczema patients and healthy controls with sufficient sensitivity and specificity. Raman spectra at different skin depths were acquired from subjects' lower volar forearm location using an in-house developed handheld confocal Raman micro-spectroscopy system. The Raman spectra corresponding to the skin surface from all the subjects were further analyzed through partial least squares discriminant analysis, a binary classification model allowing the classification between eczema and healthy subjects with a sensitivity and specificity of 0.94 and 0.85, respectively, using stratified K-fold (K = 10) cross-validation. The variable importance in the projection score from the partial least squares discriminant analysis classification model further elucidated the role of important stratum corneum proteins and lipids in distinguishing two subject groups.

Optical methods for blood perfusion measurement--theoretical comparison among four different modalities.

Blood perfusion in human tissue can be measured in vivo by means of various optical methods, which seem to be very different from one another. The most prominent examples of them are laser Doppler flowmetry, laser speckle contrast imaging, diffuse correlation spectroscopy, and the most recently developed diffuse speckle contrast analysis. In this paper, we claim that these four seemingly different modalities are examining different aspects of the same entity-the temporal autocorrelation function of scattered photons. We will show how the observables in each modality can be theoretically derived from the temporal autocorrelation function, and will discuss the merits and drawbacks of each modality in its practical use.

Hybrid Photoacoustic Ultrasound Imaging System for Cold-Induced Vasoconstriction and Vasodilation Monitoring.

Lewis hunting reaction refers to the alternating cold-induced vasoconstriction and dilation in extremities, whose underlying mechanism is complex. While numerous studies reported this intriguing phenomenon by measuring cutaneous temperature fluctuation under cold exposure, few of them tracked peripheral vascular responses in real-time, lacking a non-invasive and quantitative imaging tool. To better monitor hunting reaction and diagnose relevant diseases, we developed a hybrid photoacoustic ultrasound (PAUS) tomography system to monitor finger vessels' dynamic response to cold, together with simultaneous temperature measurement. We also came out a standard workflow for image analysis with self-defined indices. In the small cohort observational study, vascular changes in the first cycle of hunting reaction were successfully captured by the image series and quantified. Time difference between vasodilation and temperature recovery was noticed and reported for the first time, thanks to the unique capability of the PAUS imaging system in real-time and continuous vascular monitoring. The developed imaging system and indices enabled more objective and quantitative monitoring of peripheral vascular activities, indicating its great potential in numerous clinical applications.

A portable optical pulsatile flowmetry demonstrates strong clinical relevance for diabetic foot perfusion assessment.

We present a robust, cost-effective (<2000 USD), and portable optical diffuse speckle pulsatile flowmetry (DSPF) device with a flexible handheld probe for deep tissue blood flow measurement in the human foot as well as a first-in-man observational clinical study using the proposed optical device for tissue ischemia assessment and peripheral artery disease (PAD) diagnosis. Blood flow in tissue is inherently pulsatile in nature. However, most conventional methods cannot measure deep tissue-level pulsatile blood flow noninvasively. The proposed optical device can measure tissue-level pulsatile blood flow ∼6 mm underneath the skin surface. A new quantitative tissue perfusion index (TPIDSPF) based on frequency domain analysis of the pulsatile blood flow waveform is defined to assess tissue ischemia status. Through a clinical study involving 66 subjects, including healthy individuals and diabetes patients with and without PAD, TPIDSPF demonstrated strong correlations of 0.720 with transcutaneous tissue partial oxygen pressure (TcPO2) and 0.652 with toe-brachial index (TBI). Moreover, among the three methods, TPIDSPF demonstrated the highest area under the curve for PAD diagnosis among diabetes patients, with a notable value of 0.941. The promising clinical results suggest that the proposed optical method has the potential to be an effective clinical tool for identifying PAD among the diabetic cohort.

Multi-channel deep tissue flowmetry based on temporal diffuse speckle contrast analysis.

Recently, diffuse speckle contrast analysis (DSCA) was introduced as a competent modality for deep tissue blood flow measurement, where the speckle contrast is calculated over spatial domain on the CCD image of diffuse reflectance. In this paper, we introduce time-domain DSCA where temporal statistics are used for speckle contrast calculation and results in the same deep tissue flow measurement. This new modality is especially suitable for multi-channel real-time flowmetry, and we demonstrate its performance on human arm during cuff occlusion test. Independent component analysis (ICA) study on multi-channel data shows promising results about underlying physiology.

Deep tissue flowmetry based on diffuse speckle contrast analysis.

Diffuse correlation spectroscopy (DCS) is an emerging modality for noninvasive deep tissue blood flow monitoring that is becoming increasingly popular; it conducts an autocorrelation analysis of fast fluctuating photon count signals from a single speckle. In this Letter, we show that the same level of deep tissue flow information can be obtained from a much simpler analysis on the spatial distribution of the speckles that is obtained by a CCD camera, which we named diffuse speckle contrast analysis (DSCA). Both the flow phantom experiment and in vivo cuff occlusion data are presented. DSCA can be considered a new optical modality that combines DCS and laser speckle contrast imaging (LSCI), which exploits simple instrumentation and analysis and yet is sensitive to deep tissue flow.

Molecular Fingerprint Detection Using Raman and Infrared Spectroscopy Technologies for Cancer Detection: A Progress Review.

Molecular vibrations play a crucial role in physical chemistry and biochemistry, and Raman and infrared spectroscopy are the two most used techniques for vibrational spectroscopy. These techniques provide unique fingerprints of the molecules in a sample, which can be used to identify the chemical bonds, functional groups, and structures of the molecules. In this review article, recent research and development activities for molecular fingerprint detection using Raman and infrared spectroscopy are discussed, with a focus on identifying specific biomolecules and studying the chemical composition of biological samples for cancer diagnosis applications. The working principle and instrumentation of each technique are also discussed for a better understanding of the analytical versatility of vibrational spectroscopy. Raman spectroscopy is an invaluable tool for studying molecules and their interactions, and its use is likely to continue to grow in the future. Research has demonstrated that Raman spectroscopy is capable of accurately diagnosing various types of cancer, making it a valuable alternative to traditional diagnostic methods such as endoscopy. Infrared spectroscopy can provide complementary information to Raman spectroscopy and detect a wide range of biomolecules at low concentrations, even in complex biological samples. The article concludes with a comparison of the techniques and insights into future directions.

Non-invasive biochemical analysis and comparison of atopic dermatitis and psoriasis skin using handheld confocal Raman spectroscopy.

A handheld non-invasive confocal Raman system (CRS) was used to evaluate the differences in skin biochemicals between atopic dermatitis (AD) and psoriasis, which are inflammatory skin conditions. Raman spectral measurements in the fingerprint and high wavenumber region were acquired using a portable in-house CRS system with excitation lasers operating at 671 and 785 nm. It was deduced that relative amount of water decreases in the following sequence of skin: healthy, psoriasis and AD. Moreover, differential trends were observed for the subclasses of ceramides such that ceramide 3 is lower in the lesional AD and psoriasis skin as compared to healthy, while ceramide 2 showed a contrasting trend of decrease in lesional AD and increase in lesional psoriasis as opposed to healthy skin. Amount of cholesterol was significantly higher in lesional psoriasis as compared to lesional AD and healthy skin. These differences can aid in an objective classification of the skin conditions and in the formulation of new disease-specific topical treatments.

Coherent backscattering cone shape depends on the beam size.

Coherent backscattering (CBS) is a beautiful physical phenomenon that takes place in a highly scattering medium, which has potential application in noninvasive optical property measurement. The current model that explains the CBS cone shape, however, assumes the incoming beam diameter is infinitely large compared to the transport length. In this paper, we evaluate the effect of a finite scalar light illumination area on the CBS cone, both theoretically and experimentally. The quantitative relationship between laser beam size and the CBS cone shape is established by using two different finite beam models (uniform top hat and Gaussian distribution). A series of experimental data with varying beam diameters is obtained for comparison with the theory. Our study shows the CBS cone shape begins to show distortion when beam size becomes submillimeter, and this effect should not be ignored in general. In biological tissue where a normal large beam CBS cone is too narrow for detection, this small beam CBS may be more advantageous for more accurate and higher resolution tissue characterization.

Applications of Optical Fiber in Label-Free Biosensors and Bioimaging: A Review.

Biosensing and bioimaging are essential in understanding biological and pathological processes in a living system, for example, in detecting and understanding certain diseases. Optical fiber has made remarkable contributions to the biosensing and bioimaging areas due to its unique advantages of compact size, immunity to electromagnetic interference, biocompatibility, fast response, etc. This review paper will present an overview of seven common types of optical fiber biosensors and optical fiber-based ultrasound detection in photoacoustic imaging (PAI) and the applications of these technologies in biosensing and bioimaging areas. Of course, there are many types of optical fiber biosensors. Still, this paper will review the most common ones: optical fiber grating, surface plasmon resonance, Sagnac interferometer, Mach-Zehnder interferometer, Michelson interferometer, Fabry-Perot Interferometer, lossy mode resonance, and surface-enhanced Raman scattering. Furthermore, different optical fiber techniques for detecting ultrasound in PAI are summarized. Finally, the main challenges and future development direction are briefly discussed.

Handheld Multifunctional Fluorescence Imager for Non-invasive Plant Phenotyping.

Fluorescence imaging has shown great potential in non-invasive plant monitoring and analysis. However, current systems have several limitations, such as bulky size, high cost, contact measurement, and lack of multifunctionality, which may hinder its applications in a wide range of settings including indoor vertical farming. Herein, we developed a compact handheld fluorescence imager enabling multipurpose plant phenotyping, such as continuous photosynthetic activity monitoring and non-destructive anthocyanin quantification. The compact imager comprises of pulse-amplitude-modulated multi-color light emitting diodes (LEDs), optimized light illumination and collection, dedicated driver circuit board, miniaturized charge-coupled device camera, and associated image analytics. Experiments conducted in drought stressed lettuce proved that the novel imager could quantitatively evaluate the plant stress by the non-invasive measurement of photosynthetic activity efficiency. Moreover, a non-invasive and fast quantification of anthocyanins in green and red Batavia lettuce leaves had excellent correlation (>84%) with conventional destructive biochemical analysis. Preliminary experimental results emphasize the high throughput monitoring capability and multifunctionality of our novel handheld fluorescence imager, indicating its tremendous potential in modern agriculture.

Biochemical "decoding" of breast ultrasound images with optoacoustic tomography fusion: First-in-human display of lipid and collagen signals on breast ultrasound.

To date, studies which utilized ultrasound (US) and optoacoustic tomography (OT) fusion (US-OT) in biochemical differentiation of malignant and benign breast conditions have relied on limited biochemical data such as oxyhaemoglobin (OH) and deoxyhaemoglobin (DH) only. There has been no data of the largest biochemical components of breast fibroglandular tissue: lipid and collagen. Here, the authors believe the ability to image collagen and lipids within the breast tissue could serve as an important milestone in breast US-OT imaging with many potential downstream clinical applications. Hence, we would like to present the first-in-human US-OT demonstration of lipid and collagen differentiation in an excised breast tissue from a 38-year-old female.

The lateral entorhinal cortex is a hub for local and global dysfunction in early Alzheimer's disease states.

Functional network activity alterations are one of the earliest hallmarks of Alzheimer's disease (AD), detected prior to amyloidosis and tauopathy. Better understanding the neuronal underpinnings of such network alterations could offer mechanistic insight into AD progression. Here, we examined a mouse model (3xTgAD mice) recapitulating this early AD stage. We found resting functional connectivity loss within ventral networks, including the entorhinal cortex, aligning with the spatial distribution of tauopathy reported in humans. Unexpectedly, in contrast to decreased connectivity at rest, 3xTgAD mice show enhanced fMRI signal within several projection areas following optogenetic activation of the entorhinal cortex. We corroborate this finding by demonstrating neuronal facilitation within ventral networks and synaptic hyperexcitability in projection targets. 3xTgAD mice, thus, reveal a dichotomic hypo-connected:resting versus hyper-responsive:active phenotype. This strong homotopy between the areas affected supports the translatability of this pathophysiological model to tau-related, early-AD deficits in humans.

Photoacoustic Tomography Appearance of Fat Necrosis: A First-in-Human Demonstration of Biochemical Signatures along with Histological Correlation.

A 50-year-old woman with no past medical history presented with a left anterior chest wall mass that was clinically soft, mobile, and non-tender. A targeted ultrasound (US) showed findings suggestive of a lipoma. However, focal "mass-like" nodules seen within the inferior portion suggested malignant transformation of a lipomatous lesion called for cross sectional imaging, such as MRI or invasive biopsy or excision for histological confirmation. A T1-weighted image demonstrated a large lipoma that has a central fat-containing region surrounded by an irregular hypointense rim in the inferior portion, confirming the benignity of the lipoma. An ultrasound-guided photoacoustic imaging (PA) of the excised specimen to derive the biochemical distribution demonstrated the "mass-like" hypoechoic regions on US as fat-containing, suggestive of benignity of lesion, rather than fat-replacing suggestive of malignancy. The case showed the potential of PA as an adjunct to US in improving the diagnostic confidence in lesion characterization.

A Portable Ultrawideband Confocal Raman Spectroscopy System with a Handheld Probe for Skin Studies.

Confocal Raman spectroscopy (CRS) has shown potential in non-invasive skin analysis. However, current CRS systems have various limitations including a narrow detection band, large size, non-flexibility, slowness, and complexity, which hinder their clinical applications. Herein, we developed a portable ultrawideband CRS system with a fiber-based handheld probe to acquire the Raman spectra in both fingerprint and high wavenumber regions in a fast and quasi-simultaneous way. Dual-wavelength excitation with a dual-passband laser cleaning filter and high-speed fiber array multiplexer was adopted instead of a specialized grating and detector to achieve instant switching between the detection regions and improve system robustness. Preliminary in vivo results demonstrated its depth profiling capability in an ultrawide detection range for stratum corneum thickness, natural moisturizing factor, and water content quantification, indicating its great potential in a wide range of clinical and cosmeceutical applications.

Biophotonic technologies for assessment of breast tumor surgical margins-A review.

Breast conserving surgery (BCS) offering similar surgical outcomes as mastectomy while retaining breast cosmesis is becoming increasingly popular for the management of early stage breast cancers. However, its association with reoperation rates of 20% to 40% following incomplete tumor removal warrants the need for a fast and accurate intraoperative surgical margin assessment tool that offers cellular, structural and molecular information of the whole specimen surface to a clinically relevant depth. Biophotonic technologies are evolving to qualify as such an intraoperative tool for clinical assessment of breast cancer surgical margins at the microscopic and macroscopic scale. Herein, we review the current research in the application of biophotonic technologies such as photoacoustic imaging, Raman spectroscopy, multimodal multiphoton imaging, diffuse optical imaging and fluorescence imaging using medically approved dyes for breast cancer detection and/or tumor subtype differentiation toward intraoperative assessment of surgical margins in BCS specimens, and possible challenges in their route to clinical translation.