ABSTRACT
@#Sulfane sulfur species in the reactive sulfur species family include hydrogen polysulfides (H2Sn, n ≥2), which play an essential role in physiological regulation and signal transduction. As a redox pair of H2S, H2Sn can be produced through oxidation or enzyme reaction and regulate protein interaction and enzyme activity.Research has revealed that H2Sn, with higher efficiency of protein S-sulfhydration than H2S, may be responsible for some physiological functions previously attributed to H2S.Therefore, real-time detection of H2Sn is crucial for studying its physiological activity and the relationship between H2S and H2Sn.Traditional detection methods, such as mass spectrometry, are not suitable for living organisms as they require tissue cell disruption.Instead, fluorescence probes are often used for in situ real-time detection due to their high sensitivity and specificity and low biological toxicity.This review summarizes the physiological regulatory activity of H2Sn, as well as the design strategy, response mechanism, fluorescence characteristics, and biological applications of H2Sn fluorescent probes based on the structure of the response group, with a prospect of the challenges and developments in this field.
ABSTRACT
Scientific and technological innovation is the most important role in driving the development of minimally invasive surgery. After more than 30 years of development, modern mini-mally invasive surgery represented by laparoscopic surgery has gradually matured. Various types of minimally invasive surgeries have been popularized, and the difficulty of surgery has changed from extreme to limit. Surgical equipments and instruments can meet the needs of most clinical operations. The future of minimally invasive surgery has reached a crossroad, and only scientific and technological innovation can promote the development of minimally invasive surgery change lanes and overtake, ushering in new development, new methods, and a new world. For innovation, the most important thing is not knowledge, but vision and ideas. A new technological revolution will inevitably bring about changes in the industry. What changes will be ushered in the operation and treatment of diseases in the future? What will be the breakthrough of minimally invasive surgery? It is worth to wait and see. The authors discuss the development direction of minimally invasive surgery based on the recent application of hot technologies in laparoscopic surgery.
ABSTRACT
Near-infrared fluorescence imaging (NIRFI) is a new noninvasive detection and diagnosis technology, with the continuous development of NIRFI technology, now widely used in the clinic, characterized by high sensitivity, high penetration, no harmful radiation and simple equipment operation. This article describes the recent applications of NIRFI in the diagnosis and treatment of breast cancer and looks at future developments and perspectives in this field.
ABSTRACT
Objective @# To investigate the application of indocyanine green in near-infrared fluorescence imaging to determine the scope of necrotic bone resection in osteoradionecrosis of the jaw and to provide a reference for clinicians@*Methods @#Eight patients with osteoradionecrosis of the jaws were enrolled. Indocyanine green was intravenously injected through the elbow vein 10 minutes before osteotomy. After conservative resection of necrotic bone lesions based on imaging results, the scope of potential dead bone resection in the area of low fluorescence intensity was gradually expanded at an initial distance of 0.3 cm. Near-infrared fluorescence imaging and fluorescence intensity determination of bone cross-section were performed before and after extended resection. Statistical differences were analyzed. All patients with osteonecrosis underwent regular follow-up to evaluate the postoperative efficacy@*Results@#Indocyanine green was injected into all 8 patients with osteoradionecrosis for near-infrared fluorescence imaging and the scans were clear; the fluorescence intensity of fresh bone wounds with an expanded mandibular resection range of (0.95 ± 0.14) cm was (226.2 ± 15.8) au, which was higher than that based on intraoperative macroscopic observation and radiological results (108.8 ± 3.4) au, (t = 20.718, P<0.001). The postoperative follow-up improvement rate of 8 patients was 87.5%.@* Conclusion @#Near-infrared fluorescence imaging with indocyanine green can assist in the successful removal of necrotic bone until fresh bleeding of the jaw wound occurs, which has important clinical value in defining the resection range of osteoradionecrosis of the jaw.
ABSTRACT
The prescription of antibiotics empirically without confirmation of an infective etiology is on the rise. Administration of appropriate antibiotics can be guided by real-time fluorescence imaging using a point-of-care device. These composite images show the presence, type and the burden of infection. The time saved by this method over microbiological testing, especially in resource-poor settings, can lead to a paradigm shift in treatment by facilitating prompt and adequate antimicrobial therapy, surgical debridement as well as follow-up. Thumbnail sketches of a series of four cases highlighting different scenarios in which a fluorescent imaging device utilizing artificial intelligence and machine learning was found useful is presented in this report.
ABSTRACT
OBJECTIVES@#Occult cervical lymph node metastasis is the most important reason for recurrence of early-stage tongue cancer and oropharyngeal cancer. Cervical sentinel lymph node (SLN) biopsy may help to identify them. Pigment dyes and radionuclide were used to label SLN. Both of them had shortage. This study aims to investigate the application and clinical value of indocyanine green fluorescence imaging in cervical SLN biopsy for patients with early-stage tongue cancer and oropharyngeal cancer.@*METHODS@#Retrospective analysis was conducted on 23 patients with early tongue cancer and oropharyngeal cancer, who received surgical treatment and used indocyanine green as a tracer to find SLN in Hunan Cancer Hospital from April to October 2021. The detection rate of SLN was calculated and the distribution of SLN in different regions of the neck was analyzed.@*RESULTS@#SLN was successfully identified in 22 of 23 patients, with a detection rate of 95.65%. Among these 22 patients, 3 patients were found to have cancer metastasis, and the rate of occult lymph node metastasis was 13.63%. No pathologically positive lymph nodes were detected in SLN-negative patients, and thus the positive predictive rate was 100%. For patients with primary lesions located in the anterior 2/3 of the tongue, the constituent ratios of SLN in neck area I, II, III, and IV were 15.15%, 71.72%, 13.13%, and 0, respectively. For patients with primary lesions located in base of the tongue, the constituent ratios of SLN in neck area I, II, III, and IV were 0, 44.44%, 44.44%, and 11.12%, respectively.@*CONCLUSIONS@#Indocyanine green fluorescence imaging has a high detection rate with accurate positive prediction in the anterior cervical SLN biopsy in patients with early-stage tongue cancer and oropharyngeal cancer. Meanwhile, it can also reflect the lymphatic drainage of tumors located at different primary sites, which has high clinical value.
Subject(s)
Humans , Sentinel Lymph Node Biopsy/methods , Indocyanine Green , Lymphatic Metastasis/pathology , Tongue Neoplasms/surgery , Retrospective Studies , Lymph Nodes/pathology , Oropharyngeal Neoplasms/surgery , TongueABSTRACT
Ubiquitin-proteasome system (UPS) plays an important role in neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). The discovery of UPS activators for anti-neurodegenerative diseases is becoming increasingly important. In this study, we aimed to identify potential UPS activators using the high-throughput screening method with the high-content fluorescence imaging system and validate the neuroprotective effect in the cell models of AD. At first, stable YFP-CL1 HT22 cells were successfully constructed by transfecting the YFP-CL1 plasmid into HT22 cells, together with G418 screening. The degradation activity of the test compounds via UPS was monitored by detecting the YFP fluorescence intensity reflected by the ubiquitin-proteasome degradation signal CL1. By employing the high-content fluorescence imaging system, together with stable YFP-CL1 HT22 cells, the UPS activators were successfully screened from our established TCM library. The representative images were captured and analyzed, and quantification of the YFP fluorescence intensity was performed by flow cytometry. Then, the neuroprotective effect of the UPS activators was investigated in pEGFP-N1-APP (APP), pRK5-EGFP-Tau P301L (Tau P301L), or pRK5-EGFP-Tau (Tau) transiently transfected HT22 cells using fluorescence imaging, flow cytometry, and Western blot. In conclusion, our study established a high-content fluorescence imaging system coupled with stable YFP-CL1 HT22 cells for the high-throughput screening of the UPS activators. Three compounds, namely salvianolic acid A (SAA), salvianolic acid B (SAB), and ellagic acid (EA), were identified to significantly decrease YFP fluorescence intensity, which suggested that these three compounds are UPS activators. The identified UPS activators were demonstrated to clear AD-related proteins, including APP, Tau, and Tau P301L. Therefore, these findings provide a novel insight into the discovery and development of anti-AD drugs.
Subject(s)
Humans , Alzheimer Disease/drug therapy , Neuroprotective Agents , Optical Imaging , Proteasome Endopeptidase Complex , UbiquitinABSTRACT
Effective supplementation of probiotics can be beneficial to intestinal health, but in situ analysis of probiotics activity has rarely been reported. In this study, by coupling fluorescein 5-isothiocyanate (FITC) and 5(6)-carboxytetramethylrhodamine N-succinimidyl ester (5(6)-TAMRA-SE) with D-lysine, two fluorescent D-amino acids (FDAAs) probes were obtained: green probe (fluorescein-D-lysine, FDL) and red probe (TAMRA-D-lysine, TDL). Then, we tried to label the three kinds of probiotics, Lactobacillus acidophilus (LA), Lactobacillus casei (LC) and Veillonella atypica (VA) in vitro. FDAAs was applied to the labeling of intestinal flora in mice, and a method was established to investigate the oral survival rate of three commonly used probiotics. All animal experiments were approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine. The results show that the two synthetic FDAAs can be non-toxic and 100% for the in vitro labeling of the three probiotics. Known by FDAAs two-step labeling of oral probiotics, the high survival rate of LA was 92.30% ± 1.67%. The survival rates of VA and LC are similar, 84.13% ± 4.06% and 82.27% ± 2.43%, respectively. This study can quickly compare the changes of colonization survival rate of different probiotics in vivo, provide theoretical support for the in situ colonization activity of probiotics in the intestine, and guide the rational drug use of clinical probiotics.
ABSTRACT
During fluorescence-guided cancer surgery, ultra-pH sensitive (UPS) fluorescent nanoprobes has multiple advantages such as real-time imaging procedures, ultra-high imaging sensitivity as well as broad tumor detection specificity. UPS nanoprobes stay at "OFF" state at higher pH and turn into "ON" state at lower pH with emission of strong fluorescence. Moreover, the transition pH points (transition pH point, pHt) can be precisely controlled by structural-based strategy. One of the previously-reported UPS nanoprobes showed good imaging effect. However, it is still not clear about the effect of pHt on cancer imaging efficiency of UPS nanoprobes and to further identify the optimal UPS. In this study, we synthesized a series of UPS nanoprobes with pHt at 4.5, 6.2, 6.6, 7.8 by adjusting the hydrophobic blocks of UPS polymers. Each nanoprobe showed excellent stability in "OFF" state by dynamic light scattering and uniform morphology observed by transmission electron microscopy. In vitro imaging characterized the ultra-pH sensitive fluorescence transition of each probe. In vivo imaging results identified two UPS nanoprobes (NP-6.2 and NP-6.6) with superior tumor imaging effect. All animal experiments in this study were approved by the Animal Ethics Committee of Peking University Health Science Center and were strictly followed by the welfare regulations of laboratory animals of Peking University Health Science Center. Therefore, this study has explored the effect of pHt on the cancer imaging efficiency of UPS nanoprobes and provides a new idea for design of the other cancer microenvironment-responsive polymers.
ABSTRACT
Objective: Local recurrence is the main cause of treatment failure in patients with oral squamous cell carcinoma (OSCC). This study was proposed to investigate the feasibility of near infrared fluorescence (NIF) via indocyanine green (ICG) for monitoring surgical marginal in operation for OSCC patients. Methods: In 35 patients with OSCC treated surgically in the Department of Oral and Maxillofacial Surgery, Nanjing University School of Medicine, from January 2019 to June 2020, ICG (0.75 mg/kg) was administered intravenously via elbow vein at (12±1) hours before surgery, and NIF was performed intraoperatively on the surgical field and the cut edge of the surgically excised specimen, and fluorescence intensity was measured for OSCC tissue and normal oral mucosa, abnormal fluorescence signals were taken and subjected to rapid cryopathological examination. Correlation between NIF tumor boundary grading and pathological tumor boundary grading was analyzed by Spearman correlation analysis. Results: Clear ICG NIF was obtained for tumor lesions in all 35 patients, with a positive rate of 100%. The fluorescence intensity of OSCC tissue was (412.73±146.56) au, which was higher than that of normal oral mucosa tissue [(279.38±82.56) au, P<0.01]. Abnormal fluorescence signals were detected at the tumor bed and the cut edge of the surgical resection specimen in 4 patients, of which 2 cases were pathologically confirmed as cancer cell residue and 2 cases as inflammatory cell infiltration. The rate of positive detection of cut margins using ICG NIF technique in OSCC was 5.7% (2/35). Twenty of the 35 OSCC patients had grade 1, 11 of grade 2, and 4 of grade 3 tumor borders revealed by NIF of surgical resection specimens, which was positively correlated with pathological tumor border (r=0.809, P<0.001). Conclusions: ICG NIF technique can effectively detect the residual cancer cells at the incision margin, which is of great clinical value in reducing local recurrence of OSCC after surgery due to intraoperative cancer residue.
Subject(s)
Humans , Carcinoma, Squamous Cell/surgery , Head and Neck Neoplasms , Indocyanine Green , Margins of Excision , Mouth Neoplasms/surgery , Neoplasm, Residual , Optical Imaging/methods , Squamous Cell Carcinoma of Head and Neck/surgeryABSTRACT
Objective:To study the glypican 3 (GPC3) fluorescent probe imagings in hepatocellular carcinoma (HCC) tissues and to determine its prognostic value in HCC patients.Methods:The data of 87 patients who were treated at the Affiliated Hospital of Southwest Medical University from January 2019 to August 2020 were retrospectively analyzed. There were 75 males and 12 females, with the age of (56.1±11.9) years. The expressions of GPC3 were measured by immunohistochemistry and by the fluorescent probe. The results obtained by these two tests were compared. Patients were followed up for recurrence after hepatectomy. Univariate and multivariate Cox regression analyses were used to analyze factors influencing recurrence-free survival.Results:Detection of the GPC3 expression by GPC3 fluorescence probe was consistent with the results obtained by immunohistochemical studies ( Kappa=0.84, P<0.001). The positive rates of GPC3 fluorescent probe was 79.3%(69/87), compared with 80.4%(70/87) by immunohistochemistry studies, with no significant difference between the two groups ( P>0.05). The patients were then divided into the low differentiation group ( n=30) and the middle high differentiation group ( n=57) by the degrees of tumor differentiation. The fluorescence intensity in the low differentiation group was 134.4(128.0, 144.7) a. u. which was significantly different from the middle high differentiation group of 84.8(0, 108.5)a.u. ( Z=-7.52, P<0.001). The median fluorescence intensity of 87 patients with HCC was 108.6 a. u.. Multivariate Cox regression analysis showed that patients with a GPC3 fluorescence intensity ≥108.6 a. u. ( HR=2.07, 95% CI: 1.21-3.53, P=0.008) had a significant increased risk of recurrence after hepatectomy. Conclusion:The expressions of GPC3 in HCC were consistent between the studies by using either a GPC3 specific fluorescent probe or immunohistochemistry studies. A GPC3 fluorescence intensity ≥108.6 a. u. was a risk factor of recurrence after hepatectomy in patients with HCC.
ABSTRACT
Objective:This meta-analysis was conducted to systematically evaluate the short-term efficacy and safety of indocyanine green fluorescence imaging guided laparoscopic liver tumor resection (FIGLTR).Methods:A systematic search was made for the literature on indocyanine green fluorescence image-guided laparoscopic hepatectomy in randomized, semi-randomized controlled trials and observational studies, and manually search published materials and conference papers in Chinese and English and trace references included in the literature. The retrieval period was up to September 2021. The quality of included studies was evaluated, then the meta-analysis was conducted using Review Manager 5.1 software.Results:Ten studies were included with 803 patients, including 341 in the FIGLTR group and 462 in the non-FIGLTR group. Meta results showed that: Compared to the traditional resection group, indocyanine green fluorescence imaging can significantly shorten the operative time ( MD=-22.61, 95% CI: -34.20--11.03, P<0.001), reduce intraoperative bleeding ( MD=-49.17, 95% CI: -84.99--13.36, P<0.01), shorter hospital stay ( MD=-0.89, 95% CI: -7.72--0.06, P<0.05), Improve the removal rate of R 0 edge ( OR=8.80, 95% CI: 1.96-39.44, P<0.05) and reduce the incidence of postoperative complications ( OR=0.55, 95% CI: 0.34-0.87, P<0.05) of laparoscopic liver tumor resection. There were no differences found in portal block time and transfusion rate. Conclusion:Indocyanine green fluorescence imaging technology provides a new way for safe and accurate laparoscopic resection of liver tumors.
ABSTRACT
Vanin-1 is an amidohydrolase that catalyses the conversion of pantetheine into the amino-thiol cysteamine and pantothenic acid (coenzyme A precursor), which plays a vital role in multiple physiological and pathological processes. In this study, an enzyme-activated near-infrared (NIR) fluorescent probe (DDAV) has been constructed for sensitively detecting Vanin-1 activity in complicated biosamples on the basis of its catalytic characteristics. DDAV exhibited a high selectivity and sensitivity toward Vanin-1 and was successfully applied to the early diagnosis of kidney injury in cisplatin-induced kidney injury model. In addition, DDAV could serve as a visual tool for in situ imaging endogenous Vanin-1 in vivo. More importantly, Enterococcus faecalis 20247 which possessed high expression of Vanin-1 was screened out from intestinal bacteria using DDAV, provided useful guidance for the rational use of NSAIDs in clinic. Finally, oleuropein as a potent natural inhibitor for Vanin-1 was discovered from herbal medicines library using a high-throughput screening method using DDAV, which held great promise for clinical therapy of inflammatory bowel disease.
ABSTRACT
Fluorescence imaging is a noninvasive and dynamic real-time imaging technique;however,it exhibits poor spatial resolution in centimeter-deep tissues because biological tissues are highly scattering media for optical radiation.The recently developed ultrasound-controlled fluorescence(UCF)imaging is a novel imaging technique that can overcome this bottleneck.Previous studies suggest that the effective contrast agent and sensitive imaging system are the two pivotal factors for generating high-resolution UCF images ex vivo and/or in vivo.Here,this review highlights the recent advances(2015-2020)in the design and synthesis of contrast agents and the improvement of imaging systems to realize high-resolution UCF imaging of deep tissues.The imaging performances of various UCF systems,including the signal-to-noise ratio,imaging resolution,and imaging depth,are specifically discussed.In addition,the challenges and prospects are highlighted.With continuously increasing research interest in this field and emerging multidisciplinary applications,UCF imaging with higher spatial resolution and larger imaging depth may be developed shortly,which is expected to have a far-reaching impact on disease surveillance and/or therapy.
ABSTRACT
@#Nerve-specific fluorescent agents can be used as nerve markers in animals to guide surgical procedures and reduce the incidence of intraoperative nerve injury.In this study, the structure of oxazine mother nucleus was modified.A series of oxazine derivative fluorescent dyes YQN-3-YQN-6 were obtained by mass spectrometry and 1H NMR, which can highlight the peripheral nerve structure of rats.Among a series of targeted fluorescent dyes, YQN-3 had emission peaks near NIR and showed highly specific nerve targeting signals in the brachial plexus and sciatic nerves 4 h after intravenous administration.In addition, YQN-3 can accurately locate and identify recurrent laryngeal nerves during thyroidectomy, thus preserving the integrity of these nerves during surgery.With its simple synthesis and low toxicity, YQW-3 can be potentially applied for clinical neural tissue imaging.
ABSTRACT
ObjectiveTo establish a simple， fast and accurate method for locating the volatile oil in Angelicae Sinensis Radix based on frozen section and fluorescence imaging technology， and to reveal the distribution and accumulation of volatile oil in the roots of this herbal medicine. MethodAngelicae Sinensis Radix was used as the research material， the best frozen section conditions for the research material were established by comparing the effects of different cryoprotectants on the quality of frozen sections of Angelicae Sinensis Radix. The suitability of Sudan Ⅲ chemical staining and fluorescence localization for positioning the volatile oil were compared according to the loss of volatile oil and the complexity of operation process. ResultA new method for evaluating the quality of frozen sections of Angelicae Sinensis Radix was established. According to the evaluation equation， it was found that the highest score was obtained when the head， body and tail positions of Angelicae Sinensis Radix were treated with 20% glycerol， 15% glycerol and 20% sucrose， respectively. There was yellowish-brown oily substance in the oil chambers of phelloderm and secondary phloem， and oil canal of the secondary xylem of Angelicae Sinensis Radix， which could be stained orange red or orange yellow by Sudan Ⅲ， and there was green spontaneous fluorescence in the same part under the fluorescence microscope. ConclusionThe relatively complete section of Angelicae Sinensis Radix can be obtained after being treated with cryoprotectant. The volatile oil exists in the oil chambers of phelloderm and secondary phloem， and oil canal of the secondary xylem of Angelicae Sinensis Radix. This study can provide reference for observation of the accumulation sites of volatile oil in other plants.
ABSTRACT
Objective:To analyse the application of indocyanine green (ICG) fluorescence contrast technique for resection of hepatoblastoma in children and to study its value in the application.Methods:The data of 13 children with pediatric hepatoblastoma who underwent open liver resection by using the ICG real-time image guided system were collected from June 2020 to October 2021 at the Children's Hospital of Nanjing Medical University. There were 10 males and 3 females. Their ages ranged from 5 days to 63 months, with a median of 22 months. The characteristics of the ICG imaging, surgical excision and postoperative pathology were analysed.Results:The tumors showed bright fluorescence in 13 patients. The border between the tumor and normal liver tissues was clear in 12 patients, and there was no difference between the background fluorescence of the liver and fluorescence of the tumor in one patient. Regular hepatic resection was performed in 6 patients and irregular hepatic resection in 7 patients under ICG fluorescence navigation. In one patient, the left half of the liver was resected, and there was no fluorescence on the transected surface of the liver remnant. The surface of the liver remnant was scattered with fluorescence showing multiple tumors, which were then locally enucleated. Twelve specimens which were fluorescently visualized were bisected with 8 specimens showing partial fluorescence visualization of the nodules within the tumor + annular visualization at the margins, and 4 patients showing annular fluorescence visualization at the tumor margins only. In 12 children, histopathological examination of the tissues at the site with circumferential imaging showed distorted, vacuolated and densely arranged hepatocytes resembling pseudo-envelope changes due to compression, but there were no tumor cells.Conclusion:The ICG fluorescence contrast technique was very effective for resection of childhood hepatoblastoma, and the ring fluorescence contrast of the tumor edge indicated the border for tumor resection. The ICG contrast plays an important role in navigating tumor resection, especially in patients with multiple tumor nodules.
ABSTRACT
The preliminary screening of oral cancer mostly depends on the experience of clinicians, The surgical margin of tumor is mostly based on physical examination and preoperative imaging examination. It lacks real-time and objective intraoperative evaluation methods. Indocyanine green (ICG), as a safe and pollution-free organic fluorescent pigments, combined with near-infrared fluorescence imaging can be applied in the screening of early oral cancer, the determination of tumor resection margins, sentinel lymph node biopsy, cervical lymph node dissection, targeted chemotherapy, and other aspects. Near-infrared fluorescence imaging may become a key link in the early diagnosis and accurate treatment for oral cancer in the future.
Subject(s)
Humans , Indocyanine Green , Lymph Nodes , Mouth Neoplasms/therapy , Optical Imaging , Sentinel Lymph Node BiopsyABSTRACT
Indocyanine green (ICG) fluorescence imaging technology has been applied in laparoscopic surgery. It is possible to highly visualize the tumor cutting edge and liver segment boundary during laparoscopic hepatectomy. Although the application of this technique in liver tumor surgery has become more and more mature, the factors affecting the quality of fluorescence imaging are still not completely clear. In this paper, we analyzed and summarized the effects of different factors such as ICG administration scheme, imaging acquisition, tumor characteristics of patients and preoperative liver function indexes on the quality of intraoperative ICG imaging, in order to provide new ideas and practical experience for clinical practice and research.
ABSTRACT
Objective:To evaluate the skin development and repair process of X-ray radiation damage in rat with non-invasive two-photon excitation fluorescence (TPEF) imaging technology in vivo. Methods:Totally 24 SD rats were randomly divided into four groups including X-ray irradiated group (25, 35 and 45 Gy) and non-irradiation control group. At different times after irradiation, the degree of skin injury was evaluated, and the pathological changes of nicotinamide adenine dinucleotide (phosphate) [NAD(P)H] and collagen fiber fluorescence signals in epidermal cells were detected in vivo by TPEF imaging technology. Results:At 10 d post-irradiation, the skin of irradiation groups showed erythema and desquamation. At 15-20 d post-irradiation, the skin of radiation groups developed progressive exudation, edema and ulcers with increasing radiation dose. On day 25, the skin began to repair in the 25 Gy group, however, the skin of other groups still had exudation and ulcers. On day 10, NAD(P)H fluorescence signal in epidermal cells of irradiation groups decreased and the fluorescence signal of collagen fibers in papillary layer and reticular layer of irradiation groups reduced, which were significantly lower than that of normal control group ( t=24.145, 28.303, 26.989, 6.654, 7.510, 7.997, P<0.05). On day 30, fluorescence signal of NAD(P)H and collagen fibers in epidermal cells and dermis began to repair, the cell from stratum granulosum, stratum spinosum, and stratum basale in the 25 Gy group showed fluorescence signal, the other groups did not show. The fluorescence signal of collagen fibers in the 25 Gy group were gradually increased in papillary layer and reticular layer, however, they were significantly lower than normal control group ( t=115.133, 17.431, P<0.05), the skin of 45 Gy group did not show fluorescence signal of collagen fibers. Conclusions:The damage and repair process of epidermal cells and dermal collagen fiber can be detected noninvasively by TPEF imaging technology after X-ray irradiation in vivo.