Non-destructive Optical Technique Research Articles

Applications of multi-parameter sensing in pharmaceutical, agriculture and mineral industries using THz spectroscopy and Low-Wavenumber Raman spectroscopy

Multi-parameter sensing using non-destructive optical spectroscopic techniques, specifically Terahertz spectroscopy and low-wavenumber Raman spectroscopy (also known as THz-Raman spectroscopy), has significantly reshaped the landscape of analytical methodologies in various industries. This comprehensive review article elucidates the transformative impact of these advanced spectroscopic methods within the fields of pharmaceutical, agriculture, and mineral industry. As technology continues to evolve, the ability to concurrently analyse multiple parameters at the molecular level has become a cornerstone for precision and efficiency in these critical sectors. In the pharmaceutical industry, THz spectroscopy has emerged as a key tool in ensuring the quality and stability of drug formulations, while low-wavenumber Raman spectroscopy complements by offering insights into chemical structure as well as composition. The agricultural sector benefits from the capabilities of these spectroscopic techniques in optimizing crop management, soil analysis, and resource utilization. Simultaneously, the mineral industry leverages THz and low-wavenumber Raman spectroscopy for accurate mineral identification, aiding in exploration, extraction, and processing. Through this review, we aim to provide a nuanced understanding of how multi-parameter sensing through THz and low-wavenumber Raman spectroscopy is driving innovation, enhancing quality control, and fostering sustainable practices across diverse industrial landscapes.

Nondestructive Imaging of Manufacturing Defects in Microarchitected Materials.

Defects in microarchitected materials exhibit a dual nature, capable of both unlocking innovative functionalities and degrading their performance. Specifically, while intentional defects are strategically introduced to customize and enhance mechanical responses, inadvertent defects stemming from manufacturing errors can disrupt the symmetries and intricate interactions within these materials. In this study, we demonstrate a nondestructive optical imaging technique that can precisely locate defects inside microscale metamaterials, as well as provide detailed insights on the specific type of defect.

Quantifying Elastic Properties of Environmental Biofilms using Optical Coherence Elastography.

Biofilms are complex biomaterials comprising a well-organized network of microbial cells encased in self-produced extracellular polymeric substances (EPS). This paper presents a detailed account of the implementation of optical coherence elastography (OCE) measurements tailored for the elastic characterization of biofilms. OCE is a non-destructive optical technique that enables the local mapping of the microstructure, morphology, and viscoelastic properties of partially transparent soft materials with high spatial and temporal resolution. We provide a comprehensive guide detailing the essential procedures for the correct implementation of this technique, along with a methodology to estimate the bulk Young's modulus of granular biofilms from the collected measurements. These consist of the system setup, data acquisition, and postprocessing. In the discussion, we delve into the underlying physics of the sensors used in OCE and explore the fundamental limitations regarding the spatial and temporal scales of OCE measurements. We conclude with potential future directions for advancing the OCE technique to facilitate elastic measurements of environmental biofilms.

An Anti-Noise-Designed Residual Phase Unwrapping Neural Network for Digital Speckle Pattern Interferometry

DSPI (Digital Speckle Pattern Interferometry) is a non-destructive optical measurement technique that obtains phase information of an object through phase unwrapping. Traditional phase unwrapping algorithms depend on the quality of the images, which demands preprocessing such as filtering and denoising. Moreover, the unwrapping time is highly influenced by the size of the images. In this study, we proposed a new deep learning-based phase unwrapping algorithm combining the residual network and U-Net network. Additionally, we incorporated an improved SSIM function as the loss function based on camera characteristics. The experimental results demonstrated that the proposed method achieved higher quality in highly noisy phase unwrapping maps compared to traditional algorithms, with SSIM values consistently above 0.98. In addition, we applied image stitching to the network to process maps of various sizes and the unwrapping time remained around 1 s even for larger images. In conclusion, our proposed network is able to achieve efficient and accurate phase unwrapping.

A novel approach for determining coconut drink adulteration by means of laser light backscattering imaging

AbstractIn this work, the simulated adulteration of coconut drink by dilution with water was investigated using laser-light backscattering (LLB) imaging. The laser vision system consisted of six low power laser modules, emitting 1 mm diameter beams at wavelengths of 532, 635, 780, 808, 850 and 1,064 nm. The backscattering images were acquired by a grey scale camera with 12 bit resolution. Eight parameters were extracted to describe the backscattering profile. The methods of linear discriminant analysis (LDA) and partial least squares (PLS) regression were performed on LLB parameters for classifying and predicting dilution level of adulterated coconut drink samples. Based on the results, LLB signals responded sensitively to adulteration. LDA results showed that adulterated samples were correctly recognized with accuracies between 60 and 100%. PLS models were able to estimate the adulteration level of samples with coefficients of determination of 0.57–0.97 in validation. This result demonstrated the potential of laser-light backscattering imaging as a rapid and non-destructive optical technique for evaluation of coconut drink adulteration.

Visualisation of drug distribution in skin using correlative optical spectroscopy and mass spectrometry imaging

A correlative methodology for label-free chemical imaging of soft tissue has been developed, combining non-linear optical spectroscopies and mass spectrometry to achieve sub-micron spatial resolution and critically improved drug detection sensitivity. The approach was applied to visualise the kinetics of drug reservoir formation within human skin following in vitro topical treatment with a commercial diclofenac gel. Non-destructive optical spectroscopic techniques, namely stimulated Raman scattering, second harmonic generation and two photon fluorescence microscopies, were used to provide chemical and structural contrast. The same tissue sections were subsequently analysed by secondary ion mass spectrometry, which offered higher sensitivity for diclofenac detection throughout the epidermis and dermis. A method was developed to combine the optical and mass spectrometric datasets using image registration techniques. The label-free, high-resolution visualisation of tissue structure coupled with sensitive chemical detection offers a powerful method for drug biodistribution studies in the skin that impact directly on topical pharmaceutical product development.

Quantitative analysis of the degree of demineralization for bleached enamel by optical coherence tomography

BackgroundTooth bleaching imparts whitening effects along with adverse effects such as increased tooth sensitivity and enamel surface changes. Herein, we employed optical coherence tomography (OCT), a nondestructive optical detection technique, for evaluation of tooth enamel after treatment with peroxide-based bleaching agents. MethodsFifteen enamel samples were bleached using 38% acidic hydrogen peroxide-based bleach, subjected to OCT scanning, and then cross-sectioned and imaged under polarized light microscopy (PLM) and transverse microradiography (TMR). OCT cross-sectional images were compared with PLM and TMR. The depth and severity of demineralization produced in the bleached enamel were measured by OCT, PLM, and TMR. Comparison between the three techniques was performed using Kruskal-Wallis H non-parametric test and Pearson correlation. ResultsIn comparison with PLM and TMR, OCT clearly detected the changes in the enamel surface after hydrogen peroxide bleaching. Significant correlations (p<0.05) were observed in lesion depth between OCT and PLM (r=0.820), OCT and TMR (r=0.822), and TMR and PLM (r=0.861). There was no statistically significant difference in demineralization depth values measured by OCT, PLM, and TMR (p>0.05). ConclusionOCT can allow real-time, non-invasive imaging of artificially bleached tooth models and automatically measure the early changes in the enamel lesion structure upon exposure to hydrogen peroxide-based bleaching agents.

Non-Destructive Monitoring of Ripening Process of the Underutilized Fruit Kadam Using Laser-Induced Fluorescence and Confocal Micro Raman Spectroscopy

Underutilized fruits like kadam are bio-reserves of numerous biochemical and minerals and have the potential to be used as food alternatives and supplements to fulfill the needs of malnourished populations. The production of quality fruits needs monitoring throughout the growth and ripening in order to build consumer and industry confidence. The nondestructive optical spectroscopic techniques have immense potential to monitor the growth and ripening indices of the fruits. Hence, this study explores the potential of laser-induced fluorescence and confocal micro Raman spectroscopy for the reliable and cost effective monitoring of growth and ripening of kadam fruits. The analysis of the laser-induced fluorescence measurements reveals that the ripening of the kadam fruits may be monitored by analyzing the fluorescence emission of chlorophyll and carotenoids. The decrease in the intensity of chlorophyll fluorescence accompanied by the increase in the intensity of the carotenoid bands is the characteristic fluorescence signature associated with the ripening of the kadam fruits. Hence, upon the ripening of the fruits, the concentration of chlorophyll decreases considerably with the accumulation of carotenoids. This fact is further complimented by confocal micro Raman measurements, which show that as the fruit ripens, the intensity of carotenoid bands increases dramatically. The observed spectral features may be used by the horticulture scientists and nutritionists to monitor the growth, maturity, and ripening of underutilized fruits like kadam in order to maximize the quality and minimize postharvest losses.

Micro-Raman spectroscopy study of blood samples from myocardial infarction patients

Acute myocardial infarction (MI) is found to be a major causative factor for global mortality and morbidity. This situation demands necessity of developing efficient and rapid diagnostic tools to detect acute MI. Raman spectroscopy is a non-destructive optical diagnostic technique, which has high potential in probing biochemical changes in clinical samples during initiation and progress of diseases. In this work, blood was taken as the sample to examine inflammation in acute MI patients using Raman spectroscopy. Ratio of Raman peak intensities that corresponds to phenylalanine (1000 cm−1) and tyrosine (825 cm−1) can facilitate indirect information about tetrahydrobiopterin (BH4) availability, which can indicate inflammatory status in patients. This ratio obtained was higher for MI patients in comparison with control subjects. The decrease in phenylalanine and tyrosine ratio (Phe-Tyr ratio) is attributed to the prognosis of standard of care (medications like antiplatelets including aspirin, statin and revascularisation) leading to inflammation reduction. Phe-Tyr ratio estimated from the Raman spectra of blood can be exploited as a reliable method to probe inflammation due to MI. The method is highly objective, require only microliters of sample and minimal sample preparation, signifying its clinical utility.

(Digital Presentation) Characterization of Passivation Dielectrics on Silicon Through Second Harmonic Generation: Effect of Fixed Charge

In applications such as image sensors or solar cells, the performance of the device can be improved thanks to the passivation of the silicon substrate using high-k dielectrics such as alumina, hafnium dioxide etc. To assess the quality of the dielectric/semiconductor interface, conventional electrical characterization techniques can be employed, but they require fabrication of dedicated test devices. Among the methods that can be directly used at wafer-level, COCOS [1] allows obtaining electrical information such as the interface states density, but the charging of the sample surface can be an issue. In this paper, we discuss an alternative sensitive and non-destructive optical technique to characterize the electrical properties of a passivation scheme: the second harmonic generation (SHG). In centrosymmetric materials, in the dipolar approximation, the bulk SHG response is zero [2], so the signal mainly contains interface-related information. In particular, when a static electric field (Edc) is present at the interface, the SHG is reinforced and a phenomenon called EFISH (electric field SHG) appears [3]. The electric field at the interface between a dielectric and a semiconductor is related to fixed charges (Qox) and/or interface states (Dit), so the SHG signal can provide these two parameters. Additionally the technique has a potential to probe small areas (within the micrometer size).In this work, the study is focused on the ability of the SHG to measure Qox. The samples used have a layer of Al2O3 of 13nm thickness on Si (100), deposited with different processes, followed or not by an annealing. The annealing step is known to activate negative charges in alumina layers, which promotes the field effect passivation and it can also modify the interface defects [4]. The samples were measured with the SHG tool from FemtoMetrix [5]. Complementary electrical characterizations were performed with the COCOS technique on the full wafer and with capacitance versus voltage (C-V) on dedicated MOS capacitors, in order to obtain Qox. The electric field at the interface can then be calculated through Gauss’ law.Figure 1a shows a typical experimental response of the SHG versus time, obtained on two different samples. The parameters Qtot and Dit were extracted through COCOS. The time-dependence of the SHG is due to the time-dependence of the electric field during the measurement because of charging/discharging effects by carriers generated under the incident laser of the SHG. The SHG signature is obviously stronger for a high Qtot, which indeed is associated with a higher electric field at the interface. Figure 1b shows (with symbols) the experimental results of SHG versus the incidence angle (AOI) for the two samples. The increase of SHG signal for the sample with higher Qtot is confirmed, regardless of the angle of incidence.A general calibration, only based on these types of measurements is not sufficient, since optical phenomena such as transmission & interferences depend on the thickness of the layers involved. In order to better anticipate the effect of these phenomena for a future calibration, we developed a home-made simulator, based on absorption in each layer, boundary conditions at each interface and which contains the “static” interface electric field between materials. Using the electric field calculated with the charge parameters extracted from the COCOS/C-V measurements, we traced the simulated SHG versus AOI (Figure 1b, lines). The tendency observed in experiments is reproduced, even though the simulations are not fully superimposed. This mismatch is probably due to the fact that some of the material parameters in the simulations (e.g. the non-linear susceptibility values) are not very well known.The full paper will show additional samples, corresponding C-V and SHG experimental and simulated data, with the aim of proposing a calibration method that would allow the direct extraction of Qox based only on the SHG measurement.AcknowledgementsThis work was supported by the French National Plan Nano2022, within the IPCEI Nanoelectronics for Europe program.[1] M. Wilson, J. Lagowski, L. Jastrzebski, A. Savtchouk, and V. Faifer, AIP Conference Proceedings, 2001, vol. 550, no. 1, pp. 220–225.[2] R. W. Boyd, Nonlinear optics. Academic press, 2020.[3] N. M. Terlinden, G. Dingemans, V. Vandalon, R. Bosch, and W. M. M. Kessels, J. Appl. Phys., vol. 115, no. 3, p. 033708, 2014.[4] D. K. Schroder. John Wiley & Sons, 2015.[5] http://femtometrix.com Figure 1

Laser Doppler Vibrometry for Evaluating the Quality of Welds in Lithium-Ion Supercells

The inspection of the quality of welds in battery packs plays an important role in ensuring safety during the manufacturing and operation of energy-storage devices in automotive vehicles during service. This research investigated the novel application of laser Doppler vibrometry, a widely used non-destructive optical technique for modal analysis, to the post-weld evaluation of micro-TIG-welded interconnections in lithium-ion supercells. The experimental modal analysis showed features in the modal models of the supercells that were unique to their welding conditions. The comparisons between the supercells showed an absence of linear correlations between the modal parameters and the welding current, as well as differences in the welding parameters obtained from the negative and positive terminals of the cylindrical cells. These findings suggested that the modal parameters of the supercells were more strongly influenced by the rigidity of the structural materials than by the localized compliance of the welded interconnections. While this investigation demonstrated a method for using laser Doppler vibrometry to distinguish between different welding conditions in lithium-ion supercells at a structural level, further development is needed to identify the weld quality of individual interconnections.

Non-destructive assessment of milk quality using pulsed UV photoacoustic, fluorescence and near FTIR spectroscopy

The work describes the application of photoacoustic (PA), fluorescence spectroscopy, and Fourier transform near-infrared spectroscopy as non-destructive optical techniques to examine the quality of milk. The amplitude of the acoustic wave was linearly proportional to the absorbed fluence. The acoustic velocity and the fluence threshold for onset of non-linearity were decreased as the fat % increased. Initially, the PA pressure was increased with fluence but it exhibited non-linearity and occurred earlier i.e. faster as the fat % was increased. The peak pressures of 120, 160, and 180 kPa were determined for 1%, 2%, and 3.5% respectively. The corresponding acoustic transient times of 0.5, 0.44, and 0.36 µs were calculated for 1%, 2%, and 3.5% milk respectively. The absorption coefficient of milk samples was determined using the pressure-fluence slope and Grüneisen constant, which increased with fat %. The bandwidths between 350–450 nm and 450–550 nm correspond to tryptophan or valine, and Methionine amino acids respectively, and the peak at ≈315 nm is thought to be due to tyrosine. The fluorescence intensity of the sample day 1 (D1-open) decreased with time more significantly due to variations in the environmental condition. The bands between 4000 and 4500 cm−1 correspond to CH-stretch, and day 4 (D4-closed) showed the highest peak amplitudes compared to the others. Combination of N–H and O–H stretch was mainly observed between 4500 and 5000 cm−1, and the bands at 4581, 4655 cm−1 in fresh sample disappeared in D1-open and D1-closed. New bands of 4717, 4792, and 4829 cm−1 were observed.

Heat transfer and pressure loss performances for additively manufactured pin fin arrays in annular channels

Additive manufacturing technology with metal powder has facilitated the production of innovative and even more complex heat sinks. The layer-additive process offers a wide range of geometries that may be exploited for advanced cooling purposes and the final identification of the best heat sink configuration has to deal with other constraints imposed by the specific application. This contribution focuses on the thermal and fluid dynamic characterization of water cooling annular channels for applications in internal combustion engine components, where also the load-bearing capacity is requested together with the cooling performances. The heat transfer inside the annular channel was enhanced by pin fins arrays that were manufactured by selective laser melting technology in AISI 316L stainless steel. Four pin fins arrays that differ in dimensions and geometries were experimentally tested over a range of mass flow rates to compare their pressure losses and heat transfer performances against those of the smooth annular channel. Furthermore, the test coupons were analysed through optical non-destructive techniques to characterize their geometrical morphology in terms of conformity with the designed model, surface roughness and waviness. In the investigated channel Reynolds number range (from 2000 to 12,000), the pin fins arrays with the largest height-to-pin diameter ratio show better thermal performances (Nu/Nu0 always greater than 2), however their heat sink efficiencies are above unity only for Re < 3000. In addition, the 45° oriented pin arrangement seems the most promising geometry due to high thermal efficiency, lighter weight and slightly lower manufacturing cost.

Time-Domain Optical Coherence Tomography System for Determining The Extinction Coefficient and Group Refractive Index of Gelatin-based Skin Phantoms

Optical Coherence Tomography (OCT) is a non-invasive, non-destructive optical imaging technique that uses a low coherence interferometer to obtain real-time cross-sectional images of samples. OCT is notably used in biomedical applications including ophthalmology and dermatology. Aside from generating cross-sectional images, axial scans can also provide additional information about its optical properties such as extinction coefficient and refractive index. This study determines the extinction coefficients and group refractive indices of gelatin-based skin phantoms using a portable time-domain (TD) – OCT system. The gelatin-based skin phantoms were fabricated with varying concentrations of titanium dioxide (TiO2), while keeping the amount of both gelatin and water constant. By changing the proportion of the gelatin powder and TiO2, skin phantoms can then be fabricated to mimic various skin conditions, both pathologic and non-pathologic. Results of the study found a positive correlation of extinction coefficient and refractive index with TiO2 concentration. Thus, increasing TiO2 concentration also increases both extinction coefficient and group refractive index. The median extinction coefficient values of the phantoms ranged from 4.29 mm−1 to 8.48 mm−1. Literature showed that the epidermis can have extinction coefficients of 1.64-7.3 mm−1. For refractive indices of the fabricated phantoms, values ranged from 1.32 to 1.48, while studies on human participants showed that human skin has refractive index values of 1.34-1.56. Based on these properties, it is feasible to fabricate phantoms simulating the optical properties of human skin.

Fraud Detection in Meat Using Hyperspectral Imaging

Fraud detection in meat is a challenging task for researchers, consumers, industries, and regulatory agencies. Traditional approaches for fraud detection are time-consuming, complicated, laborious, and expensive; they require technical skills. Therefore, much effort has been devoted in academia and industry to developing rapid and nondestructive optical techniques for fraud detection in meat. Among them, hyperspectral imaging has gained enormous attention and curiosity throughout the world. Hyperspectral imaging is an emerging analytical technique that combines spectroscopy and imaging in one system to acquire spectra and spatial information from an object simultaneously. Hyperspectral imaging is the only analytical technology that answers commonly asked analytical questions such as what chemical species are in the samples, how much, and most importantly, where they are located. Therefore, the technology will undoubtedly play indispensable roles in research and industry for fraud detection in the coming days.

Interferometric in-situ III/V semiconductor dry-etch depth-control with ±0.8 nm best accuracy using a quadruple-Vernier-scale measurement

Semiconductor multilayer and device fabrication is a complex task in electronics and opto-electronics. Layer dry etching is one of the process steps to achieve a specific lateral device design. In situ and real-time monitoring of etch depth will be necessary if high precision in etch depth is required. Nondestructive optical techniques are the methods of choice. Reflectance anisotropy spectroscopy equipment has been used to monitor the accurate etch depth during reactive ion etching of III/V semiconductor samples in situ and real time. For this purpose, temporal Fabry–Perot oscillations due to the etch-related shrinking thickness of the uppermost layer have been exploited. Earlier, we have already reported an etch-depth resolution of ±16.0 nm. By the use of a quadruple-Vernier-scale measurement and an evaluation protocol, now we even improve the in situ real-time etch-depth resolution by a factor of 20, i.e., nominally down to ±0.8 nm.

Research advancements in optical imaging and spectroscopic techniques for nondestructive detection of mold infection and mycotoxins in cereal grains and nuts.

Cereal grains and nuts are represented as the economic backbone of many developed and developing countries. Kernels of cereal grains and nuts are prone to mold infection under high relative humidity and suitable temperature conditions in the field as well as storage conditions. Health risks caused by molds and their molecular metabolite mycotoxins are, therefore, important topics to investigate. Strict regulations have been developed by international trade regulatory bodies for the detection of mold growth and mycotoxin contamination across the food chain starting from the harvest to storage and consumption. Molds and aflatoxins are not evenly distributed over the bulk of grains, thus appropriate sampling for detection and quantification is crucial. Existing reference methods for mold and mycotoxin detection are destructive in nature as well as involve skilled labor and hazardous chemicals. Also, these methods cannot be used for inline sorting of the infected kernels. Thus, analytical methods have been extensively researched to develop the one that is more practical to be used in commercial detection and sorting processes. Among various analytical techniques, optical imaging and spectroscopic techniques are attracting growers' attention for their potential of nondestructive and rapid inline identification and quantification of molds and mycotoxins in various food products. This review summarizes the recent application of rapid and nondestructive optical imaging and spectroscopic techniques, including digital color imaging, X-ray imaging, near-infrared spectroscopy, fluorescent, multispectral, and hyperspectral imaging. Advance chemometric techniques to identify very low-level mold growth and mycotoxin contamination are also discussed. Benefits, limitations, and challenges of deploying these techniques in practice are also presented in this paper.

RAMAN SPECTROSCOPY FOR INTRAOPERATIVE ASSESSMENT OF BONE RESECTION MARGINS IN ORAL CAVITY SQUAMOUS CELL CARCINOMA

<h3>Background</h3> Oral cavity squamous cell carcinoma (OCSCC) is the most frequent head and neck cancer. Surgery is the mainstay treatment for patients with OCSCC. In case of bone involvement, the affected bone needs to be resected. Cancer-free bone resection margins (BRMs) are of crucial importance: patients with cancer-free BRMs have a 2 times higher chance of survival. Currently, there is no standard method for intraoperative assessment of BRMs. Bone margin status is only known after tissue decalcification, which takes 1 to 2 weeks. After that time reoperations are highly undesirable, because the surgical defect has healed. Therefore, it is crucial to achieve tumor-free resection surfaces, which requires the possibility of intraoperative assessment of BRMs. <h3>Objective</h3> The aim of this study was to investigate the potential of Raman spectroscopy (RS) for detection of OCSCC in bone resection surfaces during mandibulectomy. RS is a nondestructive objective optical technique that provides information about the molecular composition of tissues. <h3>Methods</h3> Raman mapping experiments were performed on fresh mandible resection specimens from patients treated with mandibulectomy for OCSCC. A tumor detection algorithm was created based on water concentration and the high-wavenumber range (2800 cm⁻¹-3050 cm⁻¹) of the Raman spectra. <h3>Results</h3> Results show that RS can detect OCSCC in bone resection surfaces with a high sensitivity (96%) and specificity (83%; 26 mapping experiments, 22 patients). <h3>Conclusions</h3> These results form the basis for further development of an RS tool as an objective method for intraoperative assessment of BRMs.

INTRAOPERATIVE ASSESSMENT OF RESECTION MARGINS BASED ON RAMAN SPECTROSCOPY IN OCSCC SURGERY

<h3>Background</h3> In head and neck oncological surgery the goal is to achieve a complete tumor resection with acceptable remaining function and appearance. For oral cavity squamous cell carcinoma (OCSCC) only 15% of the resections are reported as adequate. Since 2013, we have performed intraoperative assessment of resection margins (IOARM) in our institute, based on palpation and visual inspection of the resected specimens by pathologist and surgeon. This has resulted in an improvement of adequate resection margins from 15% to 50%, underlining the importance of IOARM. However, this method is subjective, labor intensive, and logistically challenging. <h3>Objective</h3> Our aim is to develop an objective method for fast and reliable IOARM based on Raman spectroscopy (RS). <h3>Methods</h3> RS is a non-destructive objective optical technique that provides information about the molecular composition of tissues. It can discriminate between healthy tissue and tumors. We developed a prototype Raman instrument employing a fiber-optic needle probe. The fiber-optic needle is driven into the OCSCC specimen, from the resection surface towards the tumor. Based on the Raman spectra collected along the insertion path, the location of the tumor border can be determined. From this the resection margin can be determined. <h3>Results</h3> First tests of the method show that the instrument accurately predicts the achieved resection margins. Per location the measurement and assessment takes 5 seconds. <h3>Conclusions</h3> This development signifies an important step towards a fast and objective IOARM. The fast measurement time enables an objective inspection of the margins achieved at a large number of locations of the resection surface.

Monitoring Changes in Biochemical and Biomechanical Properties of Collagenous Tissues Using Label-Free and Nondestructive Optical Imaging Techniques

We demonstrate the ability of nondestructive optical imaging techniques such as second-harmonic generation (SHG), two-photon fluorescence (TPF), fluorescence lifetime imaging (FLIM), and Raman spectroscopy (RS) to monitor biochemical and mechanical alterations in tissues upon collagen degradation. Decellularized equine pericardium (EP) was treated with 50 μg/mL bacterial collagenase at 37 °C for 8, 16, 24, and 32 h. The SHG ratio (defined as the normalized ratio between SHG and TPF signals) remained unchanged for untreated EP (stored in phosphate-buffered solution (PBS)), whereas treated EP showed a trend of a decreasing SHG ratio with increasing collagen degradation. In the fluorescence domain, treated EP experienced a red-shifted emission and the fluorescence lifetime had a trend of decreasing lifetime with increasing collagen digestion. RS monitors collagen degradation, the spectra had less intense Raman bands at 814, 852, 938, 1242, and 1270 cm-1. Non-negative least-squares (NNLS) modeling quantifies collagen loss and relative increase of elastin. The Young's modulus, derived from atomic force microscope-based nanoindentation experiments, showed a rapid decrease within the first 8 h of collagen degradation, whereas more gradual changes were observed for optical modalities. We conclude that optical imaging techniques like SHG, RS, and FLIM can monitor collagen degradation in a label-free manner and coarsely access mechanical properties in a nondestructive manner.