Objective Perfusion Assessment in Gracilis Muscle Interposition—A Novel Software-Based Approach to Indocyanine Green Derived Near-Infrared Fluorescence in Reconstructive Surgery

Background: Gracilis muscle interposition (GMI) is an established treatment option for complex perineal fistulas and reconstruction. The outcome is limited by complications such as necrosis, impaired wound healing and fistula persistence or recurrence. Quantifiable methods of assessing muscle flap perfusion intraoperatively are lacking. This study evaluates a novel and objective software-based assessment of indocyanine green near-infrared fluorescence (ICG-NIRF) in GMI. Methods: Intraoperative ICG-NIRF visualization data of five patients with inflammatory bowel disease (IBD) undergoing GMI for perineal fistula and reconstruction were analyzed retrospectively. A new software was utilized to generate perfusion curves for the specific regions of interest (ROIs) of each GMI by depicting the fluorescence intensity over time. Additionally, a pixel-to-pixel and perfusion zone analysis were performed. The findings were correlated with the clinical outcome. Results: Four patients underwent GMI without postoperative complications within 3 months. The novel perfusion indicators identified here (shape of the perfusion curve, maximum slope value, distribution and range) indicated adequate perfusion. In one patient, GMI failed. In this case, the perfusion indicators suggested impaired perfusion. Conclusions: We present a novel, software-based approach for ICG-NIRF perfusion assessment, identifying previously unknown objective indicators of muscle flap perfusion. Ready for intraoperative real-time use, this method has considerable potential to optimize GMI surgery in the future

Innovative Benthic Lander for Macroalgae Monitoring in Shallow-Water Environments

The colonization of underwater environments by exotic seaweeds is causing major ecological problems around the world. This project, referred to AMALIA, aims to transform this current ocean threat into an opportunity by adding value to the macroalgae present off the northwest of the Iberian Peninsula. To do so and to observe the presence of seaweeds in situ, an ocean modular submersible platform was developed. This platform was designed to be capable of detecting and surveying surges of invasive seaweeds while withstanding sea conditions. Conceptual designs followed by a screening process were performed, taking into consideration criteria such as operational range and modularity. An open-frame lander was considered and further developed using buckling criteria. In parallel, a state-of-the-art monitoring system was created using spectral imaging, allowing for the future creation of a macroalgae identification system. In addition, sensorial systems for characterizing growth conditions were introduced. Laboratory trials were executed to assess the capability of the system, and sea trials are currently being performed. Numerical simulations and laboratory trials indicate that the structure is fully capable of being deployed for shallow-water environments with a state-of-the-art invasive seaweed monitoring system while maintaining a high degree of modularity

Infrared imaging of the crime scene: possibilities and pitfalls

All objects radiate infrared energy invisible to the human eye, which can be imaged by infrared cameras, visualizing differences in temperature and/or emissivity of objects. Infrared imaging is an emerging technique for forensic investigators. The rapid, nondestructive, and noncontact features of infrared imaging indicate its suitability for many forensic applications, ranging from the estimation of time of death to the detection of blood stains on dark backgrounds. This paper provides an overview of the principles and instrumentation involved in infrared imaging. Difficulties concerning the image interpretation due to different radiation sources and different emissivity values within a scene are addressed. Finally, reported forensic applications are reviewed and supported by practical illustrations. When introduced in forensic casework, infrared imaging can help investigators to detect, to visualize, and to identify useful evidence nondestructivel

Optimizing Indocyanine Green Dosage for Near-Infrared Fluorescence Perfusion Assessment in Bowel Anastomosis: A Prospective, Systematic Dose-Ranging Study

Background: Indocyanine green (ICG) near-infrared fluorescence (NIRF) has emerged as a promising technique for visualizing tissue perfusion. However, within the wide range of dosages and imaging conditions currently being applied, the optimal dosage of ICG remains unclear. This study aimed to investigate the feasibility and implications of implementing lower dosages of ICG than commonly used for visual and quantitative perfusion assessment in a standardized setting. Methods: A prospective single-center cohort study was conducted on patients undergoing ileostomy reversal by hand-sewn anastomosis. ICG-NIRF visualization was performed before (T1) and after (T2) anastomosis with one of four different dosages of ICG (5 mg, 2.5 mg, 1.25 mg, or 0.625 mg) and recorded. Postoperatively, each visualization was evaluated for signal strength, completeness, and homogeneity of fluorescence. Additionally, perfusion graphs were generated by a software-based quantitative perfusion assessment, allowing an analysis of perfusion parameters. Statistical analysis comparing the effect of the investigated dosages on these parameters was performed. Results: In total, 40 patients were investigated. Visual evaluation demonstrated strong, complete, and homogeneous fluorescence signals across all dosages. Perfusion graph assessment revealed a consistent shape for all dosages (ingress followed by egress phase). While the average signal intensity decreased with dosage, it was sufficient to enable perfusion assessment even at the lowest dosages of 1.25 mg and 0.625 mg of ICG. The baseline intensity at T2 (the second intraoperative visualization) significantly decreased with dosage. The slope of the egress phase steepened with decreasing dosage. Conclusions: Lower dosages of ICG were sufficient for intraoperative perfusion assessment, while causing lower residual fluorescence and quicker egress in subsequent visualizations

Methylene Blue Near-Infrared Fluorescence Imaging in Breast Cancer Sentinel Node Biopsy

Introduction: Fluorescence-based navigation for breast cancer sentinel node biopsy is a novel method that uses indocyanine green as a fluorophore. However, methylene blue (MB) also has some fluorescent properties. This study is the first in a clinical series presenting the possible use of MB as a fluorescent dye for the identification of sentinel nodes in breast sentinel node biopsy. Material and methods: Forty-nine patients with breast cancer who underwent sentinel node biopsy procedures were enrolled in the study. All patients underwent standard simultaneous injection of nanocolloid and MB. We visualized and assessed the sentinel nodes and the lymphatic channels transcutaneously, with and without fluorescence, and calculated the signal-to-background ratio (SBR). We also analyzed the corresponding fluorescence intensity of various dilutions of MB. Results: In twenty-three patients (46.9%), the location of the sentinel node, or the end of the lymphatic path, was visible transcutaneously. The median SBR for transcutaneous sentinel node location was 1.69 (range 1.66–4.35). Lymphatic channels were visible under fluorescence in 14 patients (28.6%) prior to visualization by the naked eye, with an average SBR of 2.01 (range 1.14–5.6). The sentinel node was visible under fluorescence in 25 patients (51%). The median SBR for sentinel node visualization with MB fluorescence was 2.54 (range 1.34–6.86). Sentinel nodes were visualized faster under fluorescence during sentinel node preparation. Factors associated with the rate of visualization included diabetes (p = 0.001), neoadjuvant chemotherapy (p = 0.003), and multifocality (p = 0.004). The best fluorescence was obtained using 40 μM (0.0128 mg/mL) MB, but we also observed a clinically relevant dilution range between 20 μM (0.0064 mg/mL) and 100 μM (0.032 mg/mL). Conclusions: For the first time, we propose the clinical usage of MB as a fluorophore for fluorescence-guided sentinel node biopsy in breast cancer patients. The quenching effect of the dye may be the reason for its poor detection rate. Our analysis of different concentrations of MB suggests a need for a detailed clinical analysis to highlight the practical usefulness of the dye

Functional imaging of the ocular fundus using an 8-band retinal multispectral imaging system

Application of functional imaging in ophthalmology requires efficient imaging techniques that can detect and quantify chromophores to visualise processes in vivo. The aim of the present study was to develop and evaluate a fast and affordable imaging system. We describe an eight-band retinal multispectral imaging (MSI) system and compare it with a hyperspectral imaging (HSI) device. Determination of blood oxygen saturation was studied as proof of principle. Reflectance of incident light is measured as 1/absorbance at different wavelengths between 440 nm and 580 nm. Both devices have incorporated optical bandpass filters in a mydriatic fundus camera. The MSI system scans the retina at eight pre-defined wavelengths specific for the spectrum of haemoglobin. The HSI system acquires a full scan from 480 to 720 nm in 5 nm steps. A simple assessment of the ratio between the absorbance peaks of oxygenated haemoglobin (HbO2 ) and reduced haemoglobin (HbR) was not suitable for generating validated oxygenation maps of the retina. However, a correction algorithm that compares the measured reflectance with reflectance spectra of fully oxygenated and fully deoxygenated blood allowed our MSI setup to estimate relative oxygen saturation at higher levels, but underestimated relative oxygen saturation at lower levels. The MSI device generated better quality images than the HSI device. It allows customisation with filter sets optimised for other chromophores of interest, and augmented with extrinsic contrast imaging agents, it has the potential for a wider range of ophthalmic molecular imaging applications

Prediction of DNA concentration in fingermarks using autofluorescence properties

During criminal investigations trace DNA samples, including fingermarks, are submitted to laboratories for short tandem repeat (STR) analysis. For most common STR analysis systems a minimum amount of input DNA is required. Upon intake by the forensic laboratory the DNA concentration is estimated using quantitative polymerase chain reaction (qPCR) analysis after which most fingermarks are excluded. To tackle the problem of unnecessary processing in the lab, our study aimed to develop a method, which is able to predict the DNA content in fingermarks directly at the crime scene. Upon excitation with a UV Crime-lite, fingermark residues have autofluorescent properties. We hypothesize that the intensity of the autofluorescence signal of the fingermark content correlates to the DNA concentration in fingermarks. In this study, 164 fingermarks were examined on their autofluorescence intensity when excited at 365 nm, the number of nucleated cells, their DNA concentration and the completeness of the STR profiles. No significant correlation was observed between the DNA concentration in fingermarks and the autofluorescence signal, indicating that a high amount of autofluorescence, thus a high amount of biomaterial, does not necessarily guarantee a higher amount of DNA. In addition, the completeness of the STR profiles did not correlate to the autofluorescence signal of fingermarks. A moderate correlation was found between the predicted DNA quantity, based on the number of nucleated cells and the DNA quantity. In summary, the autofluorescence signal of fingermarks cannot directly be used as a guide to select fingermarks for DNA analysis directly at the crime scene. However, predicting the amount of DNA using a sensitive and specific DNA staining method can probably be used to estimate the DNA concentration in touch samples

Feasibility of Novel Software-Based Perfusion Indicators for the Ileal J-Pouch—On the Path towards Objective and Quantifiable Intraoperative Perfusion Assessment with Indocyanine Green Near-Infrared Fluorescence

Background: In restorative proctocolectomy with ileal J-pouch, perfusion assessment is vital to prevent complications such as anastomotic leak (AL). Indocyanine green near-infrared fluorescence (ICG-NIRF) is gaining popularity, while its interpretation and relevance remain subjective. This study aimed to evaluate a standardized ICG-NIRF imaging protocol combined with a novel, software-based assessment to detect areas of impaired perfusion and a possible correlation with AL of the pouch. Methods: In this prospective study, patients undergoing ileal J-pouch for ulcerative colitis at an inflammatory bowel disease (IBD) referral center were included. Intraoperatively, strictly standardized ICG-NIRF visualization was performed and video-recorded. Postoperatively, a specific software was utilized to determine the change in fluorescence intensity per second (i/s) for systematic regions of interest, generating perfusion-time curves and a pixel-to-pixel map. These were analysed in detail and correlated with clinical outcome (primary end point: AL within 30 days; clearly defined and screened for by pouchoscopy). Results: Four out of 18 included patients developed AL of the ileal pouch-anal anastomosis (IPAA). In the AL group, the perfusion curves on the area adjacent to the IPAA (pouch apex) displayed considerably lower ingress/inflow (median = 1.7; range = 8.5; interquartile-range = 3.8 i/s) and egress/outflow (median = −0.1; range = 0.7; interquartile-range = 0.5 i/s) values than in the non-AL group (ingress: median = 4.3; range = 10.3; interquartile-range = 4.0 i/s); egress: median = (−1.1); range = 3.9; interquartile range = 1.0 i/s). This was confirmed by further novel parameters of pouch perfusion (maximum ingress; maximum egress) and pixel-to-pixel analysis. Conclusions: This study presents the feasibility of a novel methodology to precisely assess pouch perfusion with ICG-NIRF, identifying comparable, quantifiable, and objective parameters to potentially detect perfusion-associated complications in surgery in real-time

Reconstructing the time since death using noninvasive thermometry and numerical analysis

The early postmortem interval (PMI), i.e., the time shortly after death, can aid in the temporal reconstruction of a suspected crime and therefore provides crucial information in forensic investigations. Currently, this information is often derived from an empirical model (Henssge's nomogram) describing posthumous body cooling under standard conditions. However, nonstandard conditions necessitate the use of subjective correction factors or preclude the use of Henssge's nomogram altogether. To address this, we developed a powerful method for early PMI reconstruction using skin thermometry in conjunction with a comprehensive thermodynamic finite-difference model, which we validated using deceased human bodies. PMIs reconstructed using this approach, on average, deviated no more than ±38 minutes from their corresponding true PMIs (which ranged from 5 to 50 hours), significantly improving on the ±3 to ±7 hours uncertainty of the gold standard. Together, these aspects render this approach a widely applicable, i.e., forensically relevant, method for thermometric early PMI reconstruction

Hyperspectral imaging for skin feature detection: Advances in markerless tracking for spine surgery

In spinal surgery, surgical navigation is an essential tool for safe intervention, including the placement of pedicle screws without injury to nerves and blood vessels. Commercially available systems typically rely on the tracking of a dynamic reference frame attached to the spine of the patient. However, the reference frame can be dislodged or obscured during the surgical procedure, resulting in loss of navigation. Hyperspectral imaging (HSI) captures a large number of spectral information bands across the electromagnetic spectrum, providing image information unseen by the human eye. We aim to exploit HSI to detect skin features in a novel methodology to track patient position in navigated spinal surgery. In our approach, we adopt two local feature detection methods, namely a conventional handcrafted local feature and a deep learning-based feature detection method, which are compared to estimate the feature displacement between different frames due to motion. To demonstrate the ability of the system in tracking skin features, we acquire hyperspectral images of the skin of 17 healthy volunteers. Deep-learned skin features are detected and localized with an average error of only 0.25 mm, outperforming the handcrafted local features with respect to the ground truth based on the use of optical markers