Near-infrared System Research Articles

Superconducting nanowire single-photon detector enhanced near-infrared II portable confocal microscopy for tissue imaging with indocyanine green.

In this Letter a novel, to our knowledge, approach for near-infrared (NIR) fluorescence portable confocal microscopy is introduced, aiming to enhance fluorescence imaging of biological samples in the NIR-II window. By integrating a superconducting nanowire single-photon detector (SNSPD) into a confocal microscopy, we have significantly leveraged the detection efficiency of the NIR-II fluorescence signal from indocyanine green (ICG), an FDA-approved dye known for its NIR-II fluorescence capabilities. The SNSPD, characterized by its extremely low dark count rate and optimized NIR system detection efficiency, enables the excitation of ICG with 1 mW and the capture of low-light fluorescence signals from deep regions (up to 512 µm). Consequently, our technique was able to produce high-resolution images of bio samples with a superior signal-to-noise ratio, making a substantial advancement in the field of fluorescence microscopy and offering a promising opportunity for future clinical study.

NIR-responsive CN-Pt-GEM hydrogel induces necroptosis and immunotherapeutic responses prevent postoperative recurrence and wound infection in lung carcinoma

BackgroundCancer recurrence following surgical resection is a major cause of treatment failure. Finding effective methods to prevent postoperative recurrence and wound infection is an important component of successful surgery. With the development of new nanotechnology, more treatment options have been provided for postoperative adjuvant therapy. This study presents an innovative hydrogel system that stimulates tumoricidal immunity after surgical resection of non-small cell lung cancer (NSCLC) and prevents cancer relapse.ResultsThe hydrogel system is based on the excellent photothermal conversion performance of single-atom platinum (CN-Pt) along with the delivery and release of the chemotherapy drug, gemcitabine (GEM). The system is coated onto the wound surface after tumor removal with subsequent near-infrared (NIR) photothermal therapy, which efficiently induces necroptosis of residual cancer cells, amplifies the levels of damage-associated molecular patterns (DAMPs), and increases the number of M1 macrophages. The significantly higher levels of phagocytic macrophages enhance tumor immunogenicity and sensitize cancer cells to CD8 + T-cell immunity to control postoperative recurrence, which has been verified using an animal model of postoperative lung cancer recurrence. The CN-Pt-GEM-hydrogel with NIR can also inhibit postoperative wound infection.ConclusionsThese findings introduce an alternative strategy for supplementing antitumor immunity in patients undergoing resection of NSCLC tumors. The CN-Pt-GEM-hydrogel with the NIR system also exhibits good biosafety and may be adaptable for clinical application in relation to tumor resection surgery, wound tissue filling, infection prevention, and recurrence prevention.

Near infrared light penetration in watermelon related to internal quality evaluation

The interaction between near infrared (NIR) light and the sample is a key factor to take into consideration when developing applications using NIR spectroscopy. The aim of this study was to assess the penetration depth of the NIR light through both intact watermelons and the red flesh of the cut fruit, using an interactance NIR system. The results showed a maximum penetration of between 9 and 11 mm in the assays carried out on intact watermelons and 11 mm in the watermelon flesh. The prediction performance of the different NIR system configurations used were compared by quantifying the SSC in intact watermelons in a non-destructive way. The results obtained for the assay in which the illuminated regions were separated by 28 mm (Distance 4) reported the best results for determination of SSC (%) obtaining an R2cv = 0.73 and RMSECV = 0.39 % and an R2p = 0.81 and RMSEP = 0.30 % when the set of data for that distance was split into calibration and internal validation sets.

Non-destructive testing of mechanical properties of solid wood panel based on partial least squares structural equation modeling transfer method

Calibration transfer between near infrared (NIR) spectrometers is a subtle issue in the chemometrics and process industry. Similar instruments may generate strongly different spectral responses, and regression models developed on a first NIR system can rarely be used with spectra collected by a second apparatus. In this work, two novel methods based on Structural Equation Modeling (SEM), called Enhanced Feature Extraction Approaches for factor analysis (EFEA-FA) and Enhanced Feature Extraction Approaches for spectral space transformation (EFEA-SST), were proposed to perform calibration transfer between NIR spectrometers. They were applied to a NIR nondestructive testing model for solid wood panels mechanical properties. Four different standardization algorithms were evaluated for transferring solid wood panels quality databases between a portable NIRS (InGaAs)-array spectrometer (NIRquest512) and a HSI Camera (SPECIM FX17). The results showed that EFEA-SST yielded the best model evaluation metrics (R2 and Root Mean Square Error of Prediction (RMSEP)) values for tensile strength (RMSEP=11.309, R2=0.865) parameters, while EFEA-FA gave the best fit for flexural strength (RMSEP=10.653, R2=0. 912). These results suggest the potential of two novel quality parameters prediction methods based on spectral databases transferred between diverse NIRS spectrometers.

Development of a Portable Near-Infrared Spectroscopy Tool for Detecting Freshness of Commercial Packaged Pork.

Real-time monitoring of meat quality requires fast, accurate, low-cost, and non-destructive analytical methods that can be used throughout the entire production chain, including the packaged product. The aim of this work was to evaluate the potential of a portable near-infrared (NIR) spectroscopy tool for the on-site detection of freshness of pork loin fillets in modified atmosphere packaging (MAP) stored on display counters. Pork loin slices were sealed in MAP trays under two proportions of O2/CO2/N2: High-Ox-MAP (30/40/30) and Low-Ox-MAP (5/20/75). Changes in pH, color, thiobarbituric acid reactive substances (TBARS), Warner−Bratzler shear force (WBSF), and microbiology (total viable counts, Enteriobacteriaceae, and lactic acid bacteria) were monitored over 15 days post-mortem at 4 °C. VIS-NIR spectra were collected from pork fillets before (through the film cover) and after opening the trays (directly on the meat surface) with a portable LABSPEC 5000 NIR system in diffuse reflectance mode (350−2500 nm). Quantitative NIR models by partial least squares regression (PLSR) showed a promising prediction ability for meat color (L*, a*, C*, and h*) and microbiological variables (R2VAL > 0.72 and RPDVAL > 2). In addition, qualitative models using PLS discriminant analysis obtained good accuracy (over 90%) for classifying pork samples as fresh (acceptable for consumption) or spoiled (not acceptable) based on their microbiological counts. VIS-NIR spectroscopy allows rapid evaluation of product quality and shelf life and could be used for on-site control of pork quality.

A novel system for analyzing indocyanine green (ICG) fluorescence spectra enables deeper lung tumor localization during thoracoscopic surgery.

BackgroundPalpation of tumors during thoracoscopic surgery remains difficult, and identification of deep-seated tumors may be impossible. This preclinical study investigated the usefulness of a novel indocyanine green (ICG) fluorescence spectroscopy system for tumor localization.MethodsICG was diluted to 5.0×10−2 mg/mL in fetal bovine serum (FBS) and mixed with silicone resin to prepare pseudo-tumors. Sponges of different densities and a porcine lung were placed on top of the pseudo-tumors, which were examined using a novel fluorescence spectroscopy system and a near-infrared (NIR) camera. Spectra were measured for different sponge and lung thicknesses, and the lung spectra were measured during both inflation and deflation.ResultsThe fluorescence spectroscopy system was able to identify tumors at depths ≥15 mm, while the NIR system was not. The spectroscopy system also detected tumors at greater depths when the density of the intervening material was lower. Depending on the density and thickness of the intervening material, the system could detect spectra as deep as 40 mm for sponges and 30 mm for lungs.ConclusionsThis new fluorescence spectroscopy system can be used to identify lung tumors up to a depth of 30 mm in experiments using pseudo-tumors and a porcine lung, which may aid in tumor identification during thoracoscopic surgery.

A portable NIR system for nondestructive assessment of SSC and firmness of Nanguo pears

An accurate, nondestructive, and low-cost measurement system was developed using a portable near-infrared (NIR) spectrometer (DLP NIRscan Nano), a Raspberry Pi board, a display, a lithium battery, and a self-made three-dimensional printed shell. NIR data were collected through two measurement modes (column and Hadamard transform) based on digital light processing. With this equipment, detection models of soluble solid content (SSC) and firmness, essential quality indicators of the fruit, were established via quantitative analysis using chemometrics and a hybrid wavelength selection strategy. The SSC and firmness prediction model established through the combination of the synergy interval partial least squares and genetic algorithm (Si-GA-PLS) showed higher prediction accuracy, with coefficient of determination of prediction (RP2) values of 0.9406 and 0.9119, respectively, and root-mean-square error of prediction (RMSEP) values of 0.1655 and 5.5003, respectively. A comparison of the model performance of different monochromator principles was also explored; they were found to be non-statistically significant differences from one another. Finally, data fusion was used to improve prediction ability. The results obtained by mid-level data fusion presented a better performance than using models based on one technique. Overall, the developed novel handheld detector exhibits potential for smart software applications with high accuracy.

Stepwise Energy Transfer: Near-Infrared Persistent Luminescence from Doped Polymeric Systems.

Organic near infrared (NIR) persistent-luminescence systems with bright and long-lived emission are highly valuable for applications in communication, imaging, and sensors. However, realizing these materials (especially lifetime over 0.1 s) is a challenge, mainly because of non-radiative quenching of their long-lived excitons. Herein, a universal strategy of stepwise Förster resonance energy transfer (FRET) for a bright NIR system with remarkable persistent luminescence (up to 0.2 s at 810nm) is presented, based on a new triphenylene-dye-doped polymer (triphenylene-2-ylboronic acid@poly(vinyl alcohol) (TP@PVA)) with a persistent blue phosphorescence of 3.29 s. This persistent NIR luminescence is demonstrated for application not only in NIR anti-counterfeiting but also NIR bioimaging with penetrating a piece of skin as thick as 2.0mm. By co-doping a red dye (such as Nile red) and an NIR dye Cyanine 7 (Cy7) into this doped PVA film, the shortage of spectral overlap between TP emission and Cy7 absorbance is successfully solved, through a stepwise FRET process involving triplet to singlet (TS)-FRET from TP to the intermediate red dye and then singlet to singlet (SS)-FRET to Cy7. It is noted that the efficiency of the upper TS-FRET is enhanced significantly by the lower SS-FRET, leading to high efficiencies for the continuous FRETs.

Evaluation of Near-Infrared Reflectance and Transflectance Sensing System for Predicting Manure Nutrients

Livestock manure is widely applied onto agriculture soil to fertilize crops and increase soil fertility. However, it is difficult to provide real-time manure nutrient data based on traditional lab analyses during application. Manure sensing using near-infrared (NIR) spectroscopy is an innovative, rapid, and cost-effective technique for inline analysis of animal manure. This study investigated a NIR sensing system with reflectance and transflectance modes to predict N speciation in dairy cow manure using a spiking method. In this study, 20 dairy cow manure samples were collected and spiked to achieve four levels of ammoniacal nitrogen (NH4-N) and organic nitrogen (Org-N) concentrations that resulted in 100 samples in each spiking group. All samples were scanned and analyzed using a NIR system with reflectance and transflectance sensor configurations. NIR calibration models were developed using partial least square regression analysis for NH4-N, Org-N, total solid (TS), ash, and particle size (PS). Coefficient of determination (R2) and root mean square error (RMSE) were selected to evaluate the models. A transflectance probe with a 1 mm path length had the best performance for analyzing manure constituents among three path lengths. Reflectance mode improved the calibration accuracy for NH4-N and Org-N, whereas transflectance mode improved the model predictability for TS, ash, and PS. Reflectance provided good prediction for NH4-N (R2 = 0.83; RMSE = 0.65 mg mL−1) and approximate predictions for Org-N (R2 = 0.66; RMSE = 1.18 mg mL−1). Transflectance was excellent for TS predictions (R2 = 0.97), and provided good quantitative predictions for ash and approximate predictions for PS. The correlations between the accuracy of NH4-N and Org-N calibration models and other manure parameters were not observed indicating the predictions of N contents were not affected by TS, ash, and PS.

Multifunctional Indocyanine Green Applications for Fluorescence-Guided Laparoscopic Colorectal Surgery.

Indocyanine green (ICG) could be applied for multiple functions such as fluorescent tumor localization, fluorescence lymph node mapping (FLNM), and intraoperative angiography in colorectal cancer surgery. With the near-infrared (NIR) systems, colonoscopic ICG tattooing can be used to define the early colorectal cancer that cannot be easily distinguished through the serosal surface. The lymphatic pathways can be visualized under the NIR system when ICG is injected through the submucosal or subserosal layer around the tumor. Intraoperative ICG angiography can be applied to find a favorable perfusion segment before the colon transection. Although all fluorescence functions are considered essential steps in image-guided surgery, it is difficult to perform multifunctional ICG applications in a single surgical procedure at once because complex protocols could interfere with each other. Therefore, we review the multifunctional ICG applications for fluorescent tumor localization, FLNM, and ICG angiography. We also discuss the optimal protocol for fluorescence-guided colorectal surgery.

Digital dynamic discrimination of primary colorectal cancer using systemic indocyanine green with near-infrared endoscopy

As indocyanine green (ICG) with near-infrared (NIR) endoscopy enhances real-time intraoperative tissue microperfusion appreciation, it may also dynamically reveal neoplasia distinctively from normal tissue especially with video software fluorescence analysis. Colorectal tumours of patients were imaged mucosally following ICG administration (0.25 mg/kg i.v.) using an endo-laparoscopic NIR system (PINPOINT Endoscopic Fluorescence System, Stryker) including immediate, continuous in situ visualization of rectal lesions transanally for up to 20 min. Spot and dynamic temporal fluorescence intensities (FI) were quantified using ImageJ (including videos at one frame/second, fps) and by a bespoke MATLAB® application that provided digitalized video tracking and signal logging at 30fps (Fluorescence Tracker App downloadable via MATLAB® file exchange). Statistical analysis of FI-time plots compared tumours (benign and malignant) against control during FI curve rise, peak and decline from apex. Early kinetic FI signal measurement delineated discriminative temporal signatures from tumours (n = 20, 9 cancers) offering rich data for analysis versus delayed spot measurement (n = 10 cancers). Malignant lesion dynamic curves peaked significantly later with a shallower gradient than normal tissue while benign lesions showed significantly greater and faster intensity drop from apex versus cancer. Automated tracker quantification efficiently expanded manual results and provided algorithmic KNN clustering. Photobleaching appeared clinically irrelevant. Analysis of a continuous stream of intraoperatively acquired early ICG fluorescence data can act as an in situ tumour-identifier with greater detail than later snapshot observation alone. Software quantification of such kinetic signatures may distinguish invasive from non-invasive neoplasia with potential for real-time in silico diagnosis.

Determination of the immunoglobulin G precipitation end-point by an intelligent near-infrared spectroscopy system

Precipitation is a key manufacturing unit during the immunoglobulin G (IgG) production, which guarantees the quality of the final product. Ethanol is usually used to purify IgG during the precipitation process, so it is important to monitor the ethanol concentration online. Near-infrared (NIR) spectroscopy is a powerful process analytical technology (PAT) which has been proved to be feasible to determine the ethanol concentration during the precipitation process. However, the NIR model is usually established based on the specific process, so a universal model is needed. And the clarity degree of solution will affect the quality of the spectra. Therefore, in this study an integrated NIR system was introduced to establish a universal NIR model which could predict the ethanol concentration online and determine the end-point of the whole process. First, a spectra acquisition device was designed and established in order to get high-quality NIR spectra. Then, a simple prepared ethanol NIR model was constructed to predict the actual manufacturing process. Finally, the end-point was determined to stop the peristaltic pump when the ethanol concentration reached 20%. The results showed that the spectra quality was good, model prediction was accurate, and process monitoring was accurate. In conclusion, all results indicated that the integrated NIR system could be used to monitor the biopharmaceutical process to help us understand the pharmaceutical process.

Mango internal defect detection based on optimal wavelength selection method using NIR spectroscopy

A non-destructive technique should be developed for performance analysis of mango fruits because the spongy tissue or internal defects could lower the quality of mango fruit and incur a lack of productivity. In this study, wavelength selection methods were proposed to identify the range of wavelengths for the classification of defected and healthy mango fruits. Feature selection methods were adopted here to achieve a significant selection of wavelengths. To measure the goodness of the model, the dataset was collected using the NIR (Near Infrared) spectroscopy with wavelength ranging from 673 nm–1900 nm. The classification was performed using Euclidean distance measure both in the original feature space and in FLD (Fisher's Linear Discriminant) transformed space. The experimental results showed that the lower range wavelength (673 nm–1100 nm) was the efficient wavelength for the detection of internal defects in mangoes. Further to express the effectiveness of the model, different feature selection techniques were investigated and found that the Fisher's criterion based technique appeared to be the best method for effective wavelength selection useful for classification of defected and healthy mango fruits. The optimal wavelengths were found in the range of 702.72 nm to 752.34 nm using Fisher's criterion with a classification accuracy of 84.5%. This study showed that NIR system is a useful technology for the automatic mango fruit assessment which has the potential to be used for internal defects in online sorting, easily distinguishable by those who do not meet minimum quality requirements.

Characterisation of frequency modulated wave of NIR photons transport in human lower forearm phantom

Free-space broadband frequency modulated near infrared (NIR) photon technique is used to model and characterise the optical behaviour in a lower forearm human phantom. NIR system measures the broadband insertion loss (IL) and the insertion phase (IP). Different penetration depth and path-related modulated photon movement is analysed. The study is divided into two stages. The first stage dedicated to performing broadband IL and IP measurements in a back-scattering method. The second stage dedicates to modelling the photon behaviour in stage 1 using a finite element simulator called as COMSOL. Results in both stages are analysed and compared using a signal processing method to identify the frequency modulated photon stamp associated with different layers and verifies the accuracy of the 3D FE modelling. Error shown to be less than 6% and results gives a clear understanding of how the modulated waves of photons behave when traveling in the human forearms phantom.

Multi-variable selection strategy based on near-infrared spectra for the rapid description of dianhong black tea quality.

The main objectives of the study are to understand and explore critical feature wavelengths of the obtained near-infrared (NIR) data relating to dianhong black tea quality categories, we propose a multi-variable selection strategy based on the variable space optimization from big to small which is the kernel idea of a variable combination of the improved genetic algorithm (IGA) and particle swarm optimization (PSO) in this study. A rapid description based on the NIR technology is implemented to assess black tea tenderness and rankings. First, 700 standard samples from dianhong black tea of seven quality classes are scanned using a NIR system. The raw spectra acquired are preprocessed by Savitzky-Golay (SG) filtering coupled with standard normal variate transformation (SNV). Then, the multi-variable selection algorithm (IGA-PSO) is applied to compare with the single method (the IGA and PSO) and search the optimal characteristic wavelengths. Finally, the identification models are developed using a decision tree (DT), partial least-squares discriminant analysis (PLS-DA), and support vector machine (SVM) based on different kernel functions combined with the effective features from the above variables screening paths for the discrimination of black tea quality. The results show that the IGA-PSO-SVM model with a radial basis function achieves the best predictive results with the correct discriminant rate (CDR) of 95.28% based on selected four characteristic variables in the prediction process. The overall results demonstrate that NIR combined with a multi-variable selection method can constitute a potential tool to understand the most important features involved in the evaluation of dianhong black tea quality helping the instrument manufacturers to achieve the development of low-cost and handheld NIR sensors.

Multispectrum Indocyanine Green Videography for Visualizing Brain Vascular Pathology

Currently, neurosurgical vascular surgery frequently uses indocyanine green (ICG)-videography (VG) to evaluate the blood flow in brain vessels. Although ICG-VG delineates intravascular ICG fluorescence as a high-intensity signal in gray-scale with dark background, it is hard to identify anatomical structures, including vasculature or surgical devices simultaneously. This report developed combination of a near-infrared (NIR) camera with particular sensitivity and an optical filter to observe the blood-flow conditions and anatomical structures. To overcome the specific issues of ICG-VG, we applied a high-sensitivity camera with a 980-nm NIR component to delineate anatomical and fluorescence images, detecting signals between 830 and 1000 nm simultaneously during operation. We used a diluted ICG phantom to evaluate fluorescence signal changes by changing wavelength of the emission light. For clinical use, we used a high-sensitivity NIR camera with a high-pass filter on a surgical microscope. The new NIR system detected signals between 770 and 1000 nm, and the lighting system illuminated objects mainly at 980-nm wavelength. Both images with the blood flow and anatomical structures were projected to the smart glasses in real time. In the phantom experiment, we found that the emission light with wide band widths (575-800 nm) evoked various intensities of ICG fluorescence. This new NIR system allowed us to observe ICG fluorescence and anatomical structures without image fusion or time-delay. The both information of anatomy and fluorescence was projected on wearable smart glasses. Furthermore, the new NIR system detected ICG-fluorescence signals for a longer duration than the original camera, which allowed us to achieve careful and detailed observation of more vasculature and fine vessels. This study proposes a new NIR system and emphasizes simultaneous observation of anatomy and fluorescence signals during operation. It paves the way for further possibilities in the development of optical systems. To understand the natural phenomena and combination of different scientific and clinical fields, it might be important to understand and combine not only fluorescence, but also natural science, optics, and background pathology. This simple system would be available for neuroendoscope and robotic surgery.

Design of Photostable, Activatable Near-Infrared Photoacoustic Probes Using Tautomeric Benziphthalocyanine as a Platform.

Near-infrared (NIR) imaging techniques have attracted significant attention for biological and medicinal applications due to the ability of NIR to penetrate deeply into tissues. However, there are very few stable, activatable molecular probes that can utilize NIR light in the wavelength range beyond 800 nm. Herein, we report a new activatable NIR system for photoacoustic imaging based on tautomeric benziphthalocyanines (BPcs). We found that the existence of a free hydroxyl group is crucial for NIR absorption of BPcs. Synthesized water-soluble hydroxy BPcs exhibited high photostability and no fluorescence, which are desirable features for photoacoustic imaging. We synthesized BPcs in which the free hydroxyl group was masked by an esterase-labile or an H2 O2 -labile group. The photoacoustic signals of these hydroxy-masked BPcs were increased upon NIR excitation at 880 nm in the presence of esterase or H2 O2 , respectively. These are rare examples of activatable probes utilizing NIR light at around 900 nm.

On-line glucose monitoring by near infrared spectroscopy during the scale up steps of mammalian cell cultivation process development

NIR spectroscopy is a non-destructive tool for in-situ, on-line bioprocess monitoring. One of its most frequent applications is the determination of metabolites during cultivation, especially glucose. Previous studies have usually investigated the applicability of Near Infrared (NIR) spectroscopy at one bioreactor scale but the effect of scale up was not explored. In this study, the complete scale up from shake flask (1 L) through 20 L, 100 L and 1000 L up to 5000 L bioreactor volume level was monitored with on-line NIR spectroscopy. The differences between runs and scales were examined using principal component analysis. The bioreactor runs were relatively similar regardless of scales but the shake flasks differed strongly from bioreactor runs. The glucose concentration throughout five 5000 L scale bioreactor runs were predicted by partial least squares regression models that were based on pre-processed spectra of bioreactor runs and combinations of them. The model that produced the lowest error of prediction (4.18 mM on a 29 mM concentration range) for all five runs in the prediction set was based on the combination of 20 L and 100 L data. This result demonstrated the capabilities and the limitations of an NIR system for glucose monitoring in mammalian cell cultivations.

Near-infrared bone densitometry: A feasibility study on distal radius measurement.

Osteoporosis, defined as decreased bone mineral density (BMD), poses patients in dangers for fracture risk and has become a major public health problem worldwide because of is associated morbidity, mortality and costs. Without doubt, early detection and timely intervention are important to successfully manage osteoporosis and its associated complications. The dual-energy x-ray absorptiometry (DXA) is the most popular and standard method to measure BMD. However, limitations including radiation exposure and availability restrict its application for osteoporosis screening among general population. In this study, we developed a simple method to detect human distal radius bone density based on near infrared (NIR) image system. Among 10 volunteers (including 5 young and 5 elderly participants) receiving bone density measurement using our NIR image system at the ultradistal part of bilateral distal radius, we demonstrated a strong correlation between the optical attenuation and BMD measured with DXA, which may facilitate predicting bone density status. We hope our potential NIR image system may open a new avenue for development of osteoporosis screening facilities and help in prevention of osteoporosis related fracture and its associated complications in the near future. Pearson's correlations between BMD values from the DXA and light intensity of NIR system.

Near-infrared Intraoperative Imaging of Thoracic Sympathetic Nerves: From Preclinical Study to Clinical Trial.

The sympathetic nervous system controls and regulates the activities of the heart and other organs. Sympathetic nervous system dysfunction leads to disease. Therefore, intraoperative real-time imaging of thoracic sympathetic nerves (ITSN) would be of great clinical significance for diagnosis and therapy. The aim of this experimental study was to evaluate the feasibility and validity of intraoperative ITSN using indocyanine green (ICG).Methods: ITSN using ICG was performed on 10 rabbits to determine its feasibility. Animals were allocated to two groups. The rabbits in one group received the same dose of ICG, but were observed at different times. The rabbits in the other group were administered different doses of ICG, but were observed at the same time. Signal to background ratio (SBR) was measured in regions of interest in all rabbits. Furthermore, fifteen consecutive patients with pulmonary nodules were intravenously injected with ICG 24 h preoperatively and underwent near-infrared (NIR) fluorescence imaging (FI) thoracoscopic surgeries between July 2015 and June 2016. A novel self-developed NIR and white-light dual-channel thoracoscope system was used. SBRs of thoracic sympathetic nerves were calculated in all patients.Results: In the preclinical study, we were able to precisely recognize each rabbit's second (T2) to fifth (T5) thoracic ganglia on both sides of the spine using ITSN with ICG. In addition, we explored the relationship between SBR and the injection time of ICG and that between SBR and the dose of ICG. Using the novel dual-channel thoracoscope system, we were able to locate the ganglia from the stellate ganglion (SG) to the sixth thoracic ganglion (T6), as well as the chains between these ganglia in all patients with a high SBR value of 3.26 (standard deviation: 0.57). The pathological results confirmed our findings.Conclusion: We were able to use ICG FI to distinguish thoracic sympathetic nerves during NIR thoracoscopic surgery. The technique may replace the rib-oriented method as standard practice for mapping the thoracic sympathetic nerves.