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.

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.

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.

Detection of cutaneous oxygen saturation using a novel snapshot hyperspectral camera: a feasibility study.

BACKGROUND: Tissue necrosis, a consequence of inadequate tissue oxygenation, is a common post-operative complication. As current surgical assessments are often limited to visual and tactile feedback, additional techniques that can aid in the interrogation of tissue viability are needed to improve patient outcomes. In this bi-institutional pilot study, the performance of a novel snapshot hyperspectral imaging camera to detect superficial cutaneous oxygen saturation (StO2) was evaluated. METHODS: Healthy human volunteers were recruited at two participating centers. Cutaneous StO2 of the forearm was determined by a snapshot hyperspectral camera on two separate study days during occlusion-reperfusion of the brachial artery and after induction of local vasodilation. To calculate the blood StO2 at each pixel in the multispectral image, spectra were selected, and fitting was performed over wavelengths ranging from 470 to 950 nm. RESULTS: Quantitative detection of physiological changes in cutaneous StO2 levels was feasible in all sixteen volunteers. A significant (P<0.001) decrease in cutaneous StO2 levels from 78.3% (SD: 15.3) at baseline to 60.6% (SD: 19.8) at the end of occlusion phase was observed, although StO2 levels returned to baseline after five minutes. Mean cutaneous StO2 values were similar in the same subjects on separate study days (Pearson R2: 0.92 and 0.77, respectively) at both centers. Local vasodilation did not yield significant changes in cutaneous StO2 values. CONCLUSIONS: This pilot study demonstrated the feasibility of a snapshot hyperspectral camera for detecting quantitative physiological changes in cutaneous StO2 in normal human volunteers, and serves as a precursor for further validation in perioperative studies.

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.

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 365nm, 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.

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 nondestructively.