Synthesis and Spectral Properties of Novel Water-soluble Near-infrared Fluorescent Indocyanines

Two fluorescent pentamethine and a squarylium indocyanines containing at least one p-carboxybenzyl group on N atoms in the heterocyclic rings were synthesized. They had good water solubility and photostability. Their maximum absorption and maximum emission were 600-700 nm in water. When it was anchored onto nanostructured TiO2 electrode, compared with in water, the squaraine showed double absorption peaks (one blue shifted and another red shifted) and absorption intensity of the red shift peak increased with the increase of the time of irradiation. The intensity of the blue one decreased simultaneously. We proposed that the presence of two electronic charge forms of squaraine anchored on the TiO2 film might be the reason.

A NEAR-INFRARED FLUORESCENT DEOXYGLUCOSE DERIVATIVE FOR OPTICAL IMAGING OF EXPERIMENTAL ARTHRITIS

The purpose of this study is to investigate whether a near-infrared fluorescence(NIRF)probe,Cy5.5-d-glucosamine(Cy5.5-2DG),can image arthritis in collagen-induced arthritic(CIA)mice.The presence of arthritis was verified by both visual examination and micro-computed tomography(MicroCT)imaging.CIA mice were imaged by a micro-positron emission tomography(MicroPET)scanner one hour after intravenous injection of 2-deoxy-2-[18F]fluoro-d-glucose([18F]FDG).After radioactivity of[18F]FDG decayed away,Cy5.5-2DG was injected into a lateral tail vein of the mice.Arthritic tissue targeting and retention of Cy5.5-2DG in CIA mice were evaluated and quantified by an optical imaging system.Inflammatory tissue in CIA mice was clearly visualized by[18F]FDG-MicroPET scan.NIRF imaging of Cy5.5-2DG in the same mice revealed that the pattern of localization of Cy5.5-2DG in the arthritic tissue was very similar to that of[18F]FDG.Quantification analysis further showed that[18F]FDG uptake in arthritic tissues at one hour post-injection(p.i.)and Cy5.5-2DG uptakes at different time points p.i.were all well correlated(r2 over 0.65).In conclusion,Cy5.5-DG can detect arthritic tissues in living mice.The good correlation between the[18F]FDG uptake and Cy5.5-2DG accumulation in the same arthritic tissue warrants further investigation of Cy5.5-2DG as an approach for assessment of anti-inflammatory treatments.

Methods to Minimize Optical Noise That Degrade Fluorescence Efficiency of Optical Probe for Near-Infrared Tracking in Surgical Environment

In the medical field, there are growing interests in applied research such as in vivo fluorescence monitoring because of excellent body transmission characteristic of the near-infrared light. However, optical noise by excitation light and illumination equipment for medical applications such as interior light, surgical light decrease efficiency of the fluorescent signal when observers such as surgeons confirm fluorescence signals in medical field. To solve these problems in medical field, we have analyzed external noise factors by effect on image realization, quantification of optical noise generation by external factors, and have suggested methods of minimize the optical noise in this paper. In case of fluorescence imaging in the operating room, it has been confirmed that fluorescent excitation light, interior light and surgical light are factors to generate optical noise. To acquire near-infrared fluorescence images and to compare fluorescence contrast under conditions of darkroom, interior light and surgical light, light emitting diodes (LEDs) sources that have peak wavelength at 740, 760 and 780 nm respectively were used as excitation light sources. In addition, short-pass filter which has transmission edge at 775 nm has been applied to minimize the optical noise in each external noise factor. By comparing contrast of each image before and after use of the short-pass filter, we confirmed that optical noise reduced 49%, 56% and 66% in external noise factors respectively.

A New Heptamethine Cyanine-Based Near-Infrared Fluorescent Probe for Divalent Copper Ions with High Selectivity

A new near-infrared fluorophore 2-(2-Aminoethyl) pyridine-tricarbocyanine (1) was rationally designed and synthe-sized as a fluorescent probe for detection of Cu2+ with high selectivity. The response of Probe 1 is based on the fluorescence quenching upon binding to Cu2+. The sensing performance of the proposed Cu2+-sensitive Probe 1 was then investigated. The probe can be applied to the quantification detection of Cu2+ with a linear concentration range covering from 4.8 × 10-7 to 1.6 × 10-4 mol/L and a detection limit of 9.3 × 10-8 mol/L. The experimental results showed that the response of 1 to Cu2+ was independent of pH in medium condition (pH 6.0-8.0), and exhibited excellent selectivity towards Cu2+ over other common metal cations.

Near-infrared fluorescence sentinel lymph node detection in gastric cancer: A pilot study

AIM: To investigate feasibility and accuracy of near-infrared fluorescence imaging using indocyanine green: nanocolloid for sentinel lymph node (SLN) detection in gastric cancer.METHODS: A prospective, single-institution, phase I feasibility trial was conducted. Patients suffering from gastric cancer and planned for gastrectomy were included. During surgery, a subserosal injection of 1.6 mL ICG:Nanocoll was administered around the tumor. NIR fluorescence imaging of the abdominal cavity was performed using the Mini-FLARE™ NIR fluorescence imaging system. Lymphatic pathways and SLNs were visualized. Of every detected SLN, the corresponding lymph node station, signal-to-background ratio and histopathological diagnosis was determined. Patients underwent standard-of-care gastrectomy. Detected SLNs outside the standard dissection planes were also resected and evaluated.RESULTS: Twenty-six patients were enrolled. Four patients were excluded because distant metastases were found during surgery or due to technical failure of the injection. In 21 of the remaining 22 patients, at least 1 SLN was detected by NIR Fluorescence imaging (mean 3.1 SLNs; range 1-6). In 8 of the 21 patients, tumor-positive LNs were found. Overall accuracy of the technique was 90% (70%-99%; 95%CI), which decreased by higher pT-stage (100%, 100%, 100%, 90%, 0% for respectively Tx, T1, T2, T3, T4 tumors). All NIR-negative SLNs were completely effaced by tumor. Mean fluorescence signal-to-background ratio of SLNs was 4.4 (range 1.4-19.8). In 8 of the 21 patients, SLNs outside the standard resection plane were identified, that contained malignant cells in 2 patients.CONCLUSION: This study shows successful use of ICG:Nanocoll as lymphatic tracer for SLN detection in gastric cancer. Moreover, tumor-containing LNs outside the standard dissection planes were identified.

Evaluation of COC183B2 antibody targeting ovarian cancer by near-infrared fluorescence imaging

Objective: To evaluate the imaging potential of a novel near-infrared(NIR) probe conjugated to COC183 B2 monoclonal antibodies(MAb) in ovarian cancer(OC).Methods: The expression of OC183 B2 antigen in OC was determined by immunohistochemical(IHC) staining using tissue microarrays with the H-score system and immunofluorescence(IF) staining of tumor cell lines.Imaging probes with the NIR fluorescent dye cyanine 7(Cy7) conjugated to COC183 B2 Mab were chemically engineered. OC183 B2-positive human OC cells(SKOV3-Luc) were injected subcutaneously into BALB/c nude mice. Bioluminescent imaging(BLI) was performed to detect tumor location and growth. COC183 B2-Cy7 at 1.1,3.3, 10, or 30 μg were used for in vivo fluorescence imaging, and phosphate-buffered saline(PBS), free Cy7 dye and mouse isotype immunoglobulin G(IgG)-Cy7(delivered at the same doses as COC183 B2-Cy7) were used as controls.Results: The expression of OC183 B2 with a high H-score was more prevalent in OC tissue than fallopian tube(FT) tissue. Among 417 OC patients, the expression of OC183 B2 was significantly correlated with the histological subtype, histological grade, residual tumor size, relapse state and survival status. IF staining demonstrated that COC183 B2 specifically expressed in SKOV3 cells but not HeLa cells. In vivo NIR fluorescence imaging indicated that COC183 B2-Cy7 was mainly distributed in the xenograft and liver with optimal tumor-to-background(T/B)ratios in the xenograft at 30 μg dose. The highest fluorescent signals in the tumor were observed at 96 h postinjection(hpi). Ex vivo fluorescence imaging revealed the fluorescent signals mainly from the tumor and liver. IHC analysis confirmed that xenografts were OC183 B2 positive.Conclusions: COC183 B2 is a good candidate for NIR fluorescence imaging and imaging-guided surgery in OC.

Advances in surgical techniques for gastric cancer:Indocyanine green and near-infrared fluorescence imaging.Is it ready for prime time?

Surgery is still the primary curative treatment for gastric cancer,which includes resection of the tumor with adequate margins and extended lymphadenectomy.In order to improve the operative results and the quality of life of patients,several endeavors have been made toward precision medicine through image-guided surgery,allowing access to real-time intraoperative anatomy and accurate tumor staging.The goal of the surgeon is to achieve a more precise,individualized,and less invasive surgery without compromising oncological efficiency and safety.In this perspective,we have demonstrated the role of indocyanine green(ICG)and near-infrared(NIR)fluorescence imaging method in gastric cancer surgery.This technique may be used to improve localization of the tumor,detection of sentinel lymph nodes(SLN),real-time lymphatic mapping,and blood flow assessment(anastomosis perfusion).

Near-infrared fluorescence guided esophageal reconstructive surgery: A systematic review

BACKGROUND After an esophagectomy, the stomach is most commonly used to restore continuity of the upper gastrointestinal tract. These esophago-gastric anastomoses are prone to serious complications such as leakage associated with high morbidity and mortality. Graft perfusion is considered to be an important predictor for anastomotic integrity. Based on the current literature we believe Indocyanine green fluorescence angiography(ICGA) is an easy assessment tool for gastric tube(GT) perfusion, and it might predict anastomotic leakage(AL).AIM To evaluate feasibility and effectiveness of ICGA in GT perfusion assessment and as a predictor of AL.METHODS This study was designed according to the PRISMA guidelines and registered in the PROSPERO database. PubMed and EMBASE were independently searched by 2 reviewers for studies presenting data on intraoperative ICGA GT perfusion assessment during esophago-gastric reconstruction after esophagectomy.Relevant outcomes such as feasibility, complications, intraoperative surgical changes based on ICGA findings, quantification attempts, anatomical data and the impact of ICGA on postoperative anastomotic complications, were collected by 2 independent researchers. The quality of the included articles was assessed based on the Methodological Index for Non-Randomized Studies. The 19 included studies presented data on 1192 esophagectomy patients, in 758 patients ICGA was used perioperative to guide esophageal reconstruction.RESULTS The 19 included studies for qualitative analyses all described ICGA as a safe and easy method to evaluate gastric graft perfusion. AL occurred in 13.8% of the entire cohort, 10% in the ICG guided group and 20.6% in the control group(P <0.001). When poorly perfused cases are excluded from the analyses, the difference in AL was even larger(AL well-perfused group 6.3% vs control group 20.5%, P <0.001). The AL rate in the group with an altered surgical plan based on the ICG image was 6.5%, similar to the well perfused group(6.3%) and significantly less than the poorly perfused group(47.8%)(P < 0.001), suggesting that the technique is able to identify and alter a potential bad outcome.CONCLUSION ICGA is a safe, feasible and promising method for perfusion assessment. The lower AL rate in the well perfused group suggest that a good fluorescent signal predicts a good outcome.

Mechanism of Dynamic Near-infrared Fluorescence Cholangiography of Extrahepatic Bile Ducts and Applications in Detecting Bile Duct Injuries Using Indocyanine Green in Animal Models

Fluorescence intraoperative cholangiography（IOC） is a potential alternative for identifying anatomical variation and preventing iatrogenic bile duct injuries by using the near-infrared probe indocyanine green（ICG）. However, the dynamic process and mechanism of fluorescence IOC have not been elucidated in previous publications. Herein, the optical properties of the complex of ICG and bile, dynamic fluorescence cholangiography and iatrogenic bile duct injuries were investigated. The emission spectrum of ICG in bile peaked at 844 nm and ICG had higher tissue penetration. Extrahepatic bile ducts could fluoresce 2 min after intravenous injection, and the fluorescence intensity reached a peak at 8 min. In addition, biliary dynamics were observed owing to ICG excretion from the bile ducts into the duodenum. Quantitative analysis indicated that ICG-guided fluorescence IOC possessed a high signal to noise ratio compared to the surrounding peripheral tissue and the portal vein. Fluorescence IOC was based on rapid uptake of circulating ICG in plasma by hepatic cells, excretion of ICG into the bile and then its interaction with protein molecules in the bile. Moreover, fluorescence IOC was sensitive to detect bile duct ligation and acute bile duct perforation using ICG in rat models. All of the results indicated that fluorescence IOC using ICG is a valid alternative for the cholangiography of extrahepatic bile ducts and has potential for measurement of biliary dynamics.

Near-infrared cholangiography with intragallbladder indocyanine green injection in minimally invasive cholecystectomy

Laparoscopic cholecystectomy(LC)remains one of the most commonly performed procedures in adult and paediatric populations.Despite the advances made in intraoperative biliary anatomy recognition,iatrogenic bile duct injuries during LC represent a fatal complication and consist an economic burden for healthcare systems.A series of methods have been proposed to prevent bile duct injury,among them the use of indocyanine green(ICG)fluorescence.The most commonly reported method of ICG injection is the intravenous administration,while literature is lacking studies investigating the direct intragallbladder ICG injection.This narrative mini-review aims to assess the potential applications,usefulness,and limitations of intragallbladder ICG fluorescence in LC.Authors screened the available international literature to identify the reports of intragallbladder ICG fluorescence imaging in minimally invasive cholecystectomy,as well as special issues regarding its use.Literature search retrieved four prospective cohort studies,three case-control studies,and one case report.In the three case-control studies selected,intragallbladder near-infrared cholangiography(NIRC)was compared with standard LC under white light,with intravenous administration of ICG for NIRC and with standard intraoperative cholangiography(IOC).In total,133 patients reported in the literature have been administered intragallbladder ICG administration for biliary mapping during LC.Literature includes several reports of intragallbladder ICG administration,but a standardized technique has not been established yet.Published data suggest that NIRC with intragallbladder ICG injection is a promising method to achieve biliary mapping,overwhelming limitations of IOC including intervention and radiation exposure,as well as the high hepatic parenchyma signal and time interval needed in intravenous ICG fluorescence.Evidence-based guidelines on the role of intragallbladder ICG fluorescence in LC require the assessment of further studies and multicenter data collection into large registries.

Tumor cell death visualization of renal cell carcinoma under the combined effect of the Gratiola officinalis extract and cyclophosphamide using fluorescent staining methods

Objective of the study:We used fluorescence imaging methods of apoptosis and necrosis in human renal carcinoma A498 tumor cells in vitro to reveal the indicated forms of cell death under the combined effect of flavonoid-containing extract of Gratiola officinalis and cytostatic(cyclo-phosphamide).Materials and methods:The dyes were propidium iodide and acridine orange,which were used in the"alive and dead"test.This test helped us to identify the total number of dead cells in the forms of necrosis and apoptosis and the number of cells in which apoptosis had started,it was characterized by the appearance of apoptotic bodies or nucleus pyknosis.Results:We found the most pronounced cytotoxic activity at the ratio of extract of Gratiola officinalis and cyclophosphamide concentrations of 1∶1.The number of living cells decreased when exposed to the ratio of extract and cytostatic concentrations of 2∶1.When the ratio of concentration of the extract relative to the cytostatic increased to 3∶1,the cytostatic activity of the extract began to appear,the total number of tumor cells decreased.The number of cells with nucleus pyknosis and the number of cells with apoptosis signs significantly increased at a 3∶1 ratio of extract and cytostatic concentrations,which confirms the presence of pro-apoptotic activity of the studied combination.This trend indicates the dependence of a certain form of cell death(apoptosis,necrosis)on the ratio of extract and cytostatic doses,and it also demon-strates the cytostatic and cytotoxic effects of this combination.Conclusion:Fluorescence methods of investigation in the"alive and dead"test allowed us to visualize the forms of cell death of human kidney carcinoma A498 by combined exposure to the fiavonoid-containing extract of Gratiola officinalis and cytostatic(cyclophosphamide)24 h after exposure.We found that the combination with a concentration ratio of the extract and cyclophosphamide of 3:1 has the greatest effectiveness due to stimulation of the cytostatic effect and cytotoxic effect.

Superiority of indocyanine green-enhanced near-infrared fluorescence-guided imaging for laparoscopic lymph node dissection in patients with early-stage endometrial cancer: A retrospective cohort study

Objective:Laparoscopic pelvic lymph node dissection(LPND),which is an effective therapy for endometrial cancer,is challenging because of the complexity of the procedure and the occurrence of postoperative complications.This study aimed to explore whether indocyanine green(ICG)-enhanced nearinfrared(NIR)fluorescence-guided LPND is superior to LPND in the context of early-stage endometrial carcinoma.Methods:In this retrospective study,we included the medical records of 190 patients with early-stage endometrioid adenocarcinoma who underwent LPND at the Department of Obstetrics and Gynecology,Sir Run Run Shaw Hospital,Zhejiang University School of Medicine between January 2019 and January 2021.Depending on whether ICG-enhanced NIR fluorescence guidance was used,the patients were assigned to the ICG group or non-ICG group.Patients were followed-up for one year after surgery.Data on demographic characteristics,pathological results,operative outcomes,and complications were collected and analyzed.Results:The baseline characteristics were comparable between the ICG group and non-ICG group,including age,BMI,pregnancy history,and preoperative hemoglobin.For surgical outcomes,the patients in ICG group had significantly lower intraoperative blood loss(50 mL vs.120 mL,p<0.001),less postoperative pelvic drainage time(4.14±1.44 d vs.5.70±1.89 d,p¼0.001),shorter duration of hospital stay(5.26±1.41 d vs.7.37±1.85 d,p¼0.003),higher number of positive pelvic lymph nodes(PLNs)(1 vs.0,p¼0.003),and more PLN-positive cases(16.0%vs.3.6%,p¼0.003)than the patients in non-ICG group.However,no significant differences were noted in blood transfusion requirement,operative time,hemoglobin level decreases,number of PLNs harvested,or the presence of lymphocysts between the two groups.Conclusion:Our study showed that ICG-enhanced NIR fluorescence-guided operation may improve the accuracy and safety of LPND.

Novel imaging system for positioning of the indocyanine green (ICG) target;visible projection of the near-infrared fluorescence image

Background: Even though NIR fluorescence imaging has many advantages in SLN mapping and cancer detection, NIR fluorescence imaging shows a serious drawback that NIR cannot be detected by the naked eye without any detectors. This limitation further disturbs accurate SLN detection and adequate tumor resection resulting in the presence of cancerous cells near the boundaries of surgically removed tissues. Materials and methods: To overcome the drawback of the conventional NIR imaging method, we suggest a novel NIR imaging system which can make the NIR fluorescence image visible to the naked eye as NIR fluorescence image detected by a video camera is processed by a computer and then projected back onto the NIR fluorescence excitation position with a projector using conspicuous color light. Image processing techniques were used for projection onto the exact position of the NIR fluorescence image. Also, we implemented a phantom experiment to evaluate the performance of the developed NIR fluorescence projection system by use of the ICG. Results: The developed NIR fluorescence projection system was applied in normal mouse model to confirm the usefulness of the system in the clinical field. A BALB/c nude mouse was prepared to be applied in normal mouse model and 0.25 mg/ml stock solution of the ICG was injected through a tail vein of the mouse. From the application in normal mouse model, we could confirm that the injected ICG stayed in the liver of the mouse and verify that the projection system projected the ICG fluorescence image at the exact location of the ICG by performing laparotomy of the mouse. Conclusions: From the application in normal mouse model, we could verify that the ICG fluorescence image was precisely projected back on the site where ICG fluorescence generated. It can be demonstrated that the NIR fluorescence projection system can make it possible to visualize the invisible NIR fluorescence image and to realize that SLN mapping and cancer detection in clinical surgery.

Constant Stress Accelerated Life Tests for Vacuum Fluorescent Display Based on Least Square Method

To estimate the life of vacuum fluorescent display （VFD） more accurately and reduce test time and cost, four constant stress accelerated life tests （CSALTs） were conducted on an accelerated life test model. In the model, statistical analysis of test data is achieved by applying lognormal function to describe the life distribution, and least square method （LSM） to calculate the mean value and the standard deviation of logarithm. As a result, the accelerated life equation was obtained, and then a self-developed software was developed to predict the VFD life. The data analysis results demonstrate that the VFD life submits to lognormal distribution, that the accelerated model meets the linear Arrhenius equation, and that the precise accelerated parameter makes it possible to acquire the life information of VFD within one month.

A New Sample-Selection and Modeling Method Based on Near-Infrared Spectroscopy and Its Industrial Application

Near-infrared( NIR) spectroscopy has been widely employed as a process analytical tool( PAT) in various fields; the most important reason for the use of this method is its ability to record spectra in real time to capture process properties. In quantitative online applications,the robustness of the established NIR model is often deteriorated by process condition variations,nonlinear of the properties or the high-dimensional of the NIR data set. To cope with such situation,a novel method based on principal component analysis( PCA) and artificial neural network( ANN) is proposed and a new sample-selection method is mentioned. The advantage of the presented approach is that it can select proper calibration samples and establish robust model effectively. The performance of the method was tested on a spectroscopic data set from a refinery process. Compared with traditional partial leastsquares( PLS),principal component regression( PCR) and several other modeling methods, the proposed approach was found to achieve good accuracy in the prediction of gasoline properties. An application of the proposed method is also reported.

Optical Probe for Near-Infrared (NIR) Fluorescence Signal Detection with High Optical Performance and Thermal Stability

We propose a new optical probe for near-infrared (NIR) fluorescence signal detection with high optical performance and thermal stability. The optical probe is composed of an optical source part for efficient excitation of NIR fluorescence signal, a heat dissipation part for stable operation of the NIR fluorescence probe, and an optical detection part for efficient detection of NIR fluorescence signal. From a simulation by use of an optical simulation tool, Light ToolsTM, we could confirm that the optical probe has optical propagation efficiency of 79.6% in case of using a circular detector with 20 cm in diameter located at 20 cm in distance from the optical source. From a measurement of temperature variation of the optical probe, we could also confirm that the optical probe has thermal stability with a standard deviation of 2.19&deg;C under room temperature condition. Finally, from an evaluation of fluorescence image quality, we could confirm that an optical noise which can bring on by overlapped band between optical spectrum of the optical source for fluorescence excitation and optical spectrum of the emitted fluorescence signal decreased effectively in the optical probe.

Determination of Anionic Surfactants in Environmental Water Samples Using Fluorescence Enhancement Method Based on Fe_3O_4@PAA-RB Fluorescent Nanoparticles

[ Objective] The study aims at developing a novel fluorescence enhancement method to determine anionic surfactants. [ Method] Based on Fe3O4 @ PAA-RB fluorescent nanoparticles as fluorescent probes, we have developed a novel fluorescence enhancement method for the determi- nation of an anionic surfactant sodium dodecyl sulfate （SDS） through the gradual optimization of experiment conditions. [ Result] Under the opti- mum conditions, the extent of fluorescence enhancement is directly proportional to SDS concentration varying from 0.5 to 16.0 μmol/L, and the de- tection limit reaches 0.051 μmol/L. The relative standard deviation （RSD） for 4.0 μmol/L SDS is 3.3% （ n =6）. The proposed method has been successfully applied to the determination of SDS in environmental water samples, with recovery of 96.3% -105.5%. E Conclusion] The novel fluo- rescence enhancement method is not only simple and rapid, but also has avoided using tedious solvent-extraction and toxic organic solvents.

ACTIVATION OF NUCLEOLAR rRNA GENE TRANSCRIPTION IN LYMPHOCYTES BY TUMOR PROMOTERS: APPLIES STUDIES WITH IN SITU HYBRIDIZATION DETECTED BY FLUORESCENT METHOD

The distribution of rDNA was visualized in interphase nuclei of tumor promoter treated human lymphocytes in comparison with the mltogen Phytoheamagglutinin (PHA) effects by using an in situ hybridization fluorescent method. The procedure involves biotinylated rDNA as the probe and FITC-avidin as detection system. Silver (Ag) staining was used to visualize nucleoli. In the interphase nuclei of most of the nonstimulated control lymphocytes, only one small Ag-stained nucleolus could be seen. The in situ hybridization, however, revealed one to several agglomerations of rDNA fluorescent spots. With tumor promoting herb extract WCE (40 μg/ml) or TPA (60 ng/ ml) treatment, the Interphase nucleoli increased slightly in number with the morphology alteration into larger, reticular or compact granular types. Ag-stained particles also increased in number. The number of the in situ hybridization rDNA fluorescent spots and dots increased markedly and largereticulate formations of numerous rDNA spots were seen. This phenomenum resembles to the changes in PHA stimulated lymphocytes. Statistic analysized data showed signifcant difference between control and drug- treated cells. These results indicate that transcriptionally activated rDNA and amplification of total rDNA was induced by the used tumor promoters.

Recent developments of fluorescence imaging technology in the second near-infrared window

Background:Fluorescence bio-imaging in the second near-infrared window(NIR-II FL,1000-1700nm)has great potential in clinical theranostics,which is of great importance providing precise locations of lesions and molecular dynamic actions simultaneously in a single nanoprobe.Methods:T here has been an upsurge of multidisciplinary research focusing on developing functional types of inorganic and organic nanoprobes that can be used for NIR-II FL with the high spatiotemporal resolution,deep tissue penetration,and negligible auto-fluorescence.Results:In this mini-review,we summarize recent progress in inorganic/organic NIR-II FL nanoprobes.We introduce the design and properties of inorganic and organic nanoprobes,in the order of single-walled carbon nanotubes,quantum dots,rare-earth-doped nanoparticles,metal nanoclusters and organic fluorophores,expect to realize precise diagnosis and efficient image-guided therapy.Conclusion:Meanwhile,to elucidate the problems and perspectives,we aim to offer diverse biological applications of inorganic/organic NIR-II FL nanoprobes and accelerate the clinical transformation progress.

应用荧光染色剂Fluorescent Brightener 28和Propidium Iodide染色识别家蚕微孢子虫

家蚕微粒子病被列为蚕种生产的唯一法定检疫对象,母蛾镜检家蚕微孢子虫(Nosema bombycis,Nb)是主要的检疫方法,但其准确性受到多种因素影响。为探究应用荧光增白剂28(Fluorescent Brightener 28,FB28)和碘化丙啶(Propidium Iodide,PI)染色识别Nb孢子的可行性,以感染Nb的家蚕幼虫中肠、蚕卵、母蛾、蚁蚕为材料,联合应用以上2种荧光染色剂进行Nb孢子检测效果试验。在荧光显微镜下可见成熟的Nb孢子被FB28染成蓝色且细胞核被PI染成红色,而未成熟的孢子只能被PI染色。采用2种荧光染色剂染色,在被感染的幼虫中肠、蚕卵、母蛾、蚁蚕中都能够检测到Nb孢子,并且带毒母蛾研磨液样品稀释1 000倍依然可以检测到Nb孢子。幼虫中肠感染后1~2 d很难观察到Nb孢子,感染后3 d可以观察到局部存在少量未成熟的Nb,感染后4 d开始局部出现成熟的Nb,至感染后5~6 d可见组织中大面积分布有成熟孢子和未成熟孢子。荧光显微镜下观察被染色的Nb孢子的细胞核明显小于幼虫中肠细胞核,且亮度明显较强,卵壳、母蛾、蚁蚕表面的几丁质碎片形状与Nb孢子明显不同,并且不能被PI染色。试验结果表明,该染色检测方法可以排除蚕体组织细胞、几丁质碎片的干扰,提高对Nb检测的准确度,并且在Nb浓度较低的情况下,可以提高检测的灵敏度。