Efficient Assessment of Tumor Vascular Shutdown by Photodynamic Therapy on Orthotopic Pancreatic Cancer Using High-Speed Wide-Field Waterproof Galvanometer Scanner Photoacoustic Microscopy.

To identify the vascular alteration by photodynamic therapy (PDT), the utilization of high-resolution, high-speed, and wide-field photoacoustic microscopy (PAM) has gained enormous interest. The rapid changes in vasculature during PDT treatment and monitoring of tumor tissue activation in the orthotopic pancreatic cancer model have received limited attention in previous studies. Here, a fully two-axes waterproof galvanometer scanner-based photoacoustic microscopy (WGS-PAM) system was developed for in vivo monitoring of dynamic variations in micro blood vessels due to PDT in an orthotopic pancreatic cancer mouse model. The photosensitizer (PS), Chlorin e6 (Ce6), was utilized to activate antitumor reactions in response to the irradiation of a 660 nm light source. Microvasculatures of angiogenesis tissue were visualized on a 40 mm2 area using the WGS-PAM system at 30 min intervals for 3 h after the PDT treatment. The decline in vascular intensity was observed at 24.5% along with a 32.4% reduction of the vascular density at 3 h post-PDT by the analysis of PAM images. The anti-vascularization effect was also identified with fluorescent imaging. Moreover, Ce6-PDT increased apoptotic and necrotic markers while decreasing vascular endothelial growth factor (VEGF) expression in MIA PaCa-2 and BxPC-3 pancreatic cancer cell lines. The approach of the WGS-PAM system shows the potential to investigate PDT effects on the mechanism of angiographic dynamics with high-resolution wide-field imaging modalities.

A depth-resolved quantitative evaluation method for non-carious cervical lesions treatment with optical coherence tomography.

OBJECTIVES: The aim of this study is prognostic assessment of surface smoothness and the presence of internal bubbles after treatment of non-cancerous cervical lesions (NCCLs) using optical coherence tomography (OCT). METHODS: After treatment with NCCLs, cross-sectional images of the lesion parts of the sample were non-invasively acquired and analyzed. The surface smoothness between tooth and resin, resin and cemento-enamel junction, and the presence bubble inside resin was confirmed. In addition, using an algorithm that distinguishes between resin and dental structure based on OCT cross-sectional images, we quantitatively analyzed the amount of resin used in treating NCCLs and acquired 3D images. RESULTS: The inner structure of the resin in each sample was checked, and the presence of bubbles was confirmed. In addition, the resin sections were separated from the tomographic images acquired by OCT to visualize 3D images. The volume of resin used in the treatment part of each NCCLs samples was quantitatively analyzed as 3.7216 ∼ 14.889 mm3. CONCLUSIONS: OCT is able to measure not only the surface abrasion provided by existing intraoral scanner, but also the size and depth location of interal bubbles, which is distinctive advantage of our method. Based on our results, OCT is a significant tool for qualitative and quantitative analysis of dental NCCLs treatment before and after treatment. CLINICAL SIGNIFICANCE: The study used OCT, a non-destructive diagnostic, to reveal the structure of the resin and the location and size of bubbles after NCCLs treatment. These findings could be golden standard in determining the prognosis of NCCLs treatment.

Integrating Simultaneous Motor Imagery and Spatial Attention for EEG-BCI Control.

OBJECTIVE: EEG-based brain-computer interfaces (BCI) are non-invasive approaches for replacing or restoring motor functions in impaired patients, and direct brain-to-device communication in the general population. Motor imagery (MI) is one of the most used BCI paradigms, but its performance varies across individuals and certain users require substantial training to develop control. In this study, we propose to integrate a MI paradigm simultaneously with a recently proposed Overt Spatial Attention (OSA) paradigm, to accomplish BCI control. METHODS: We evaluated a cohort of 25 human subjects' ability to control a virtual cursor in one- and two-dimensions over 5 BCI sessions. The subjects used 5 different BCI paradigms: MI alone, OSA alone, MI, and OSA simultaneously towards the same target (MI+OSA), and MI for one axis while OSA controls the other (MI/OSA and OSA/MI). RESULTS: Our results show that MI+OSA reached the highest average online performance in 2D tasks at 49% Percent Valid Correct (PVC), and statistically outperforms both MI alone (42%) and OSA alone (45%). MI+OSA had a similar performance to each subject's best individual method between MI alone and OSA alone (50%) and 9 subjects reached their highest average BCI performance using MI+OSA. CONCLUSION: Integrating MI and OSA leads to improved performance over both individual methods at the group level and is the best BCI paradigm option for some subjects. SIGNIFICANCE: This work proposes a new BCI control paradigm that integrates two existing paradigms and demonstrates its value by showing that it can improve users' BCI performance.

A concept to detect a subgingival finish line using an intraoral optical coherence tomography system: A clinical report.

This clinical report introduces an approach for detecting the supragingival finish line by penetrating the teeth and gingival tissue using optical coherence tomography (OCT) technology. This approach was used in 3 patients who underwent tooth preparation with a subgingival finish line. Consequently, the subgingival finish line, typically challenging to discern clearly in intraoral scans, was identifiable in the OCT image.

Skin pore imaging using spectral-domain optical coherence tomography: a case report.

Sebum is an important component of the skin that has attracted attention in many fields, including dermatology and cosmetics. Pore expansion due to sebum on the skin can lead to various problems. Therefore, it is necessary to analyze the morphological characteristics of sebum. In this study, we used optical coherence tomography (OCT) to evaluate facial sebum areas. We obtained the OCT maximum amplitude projection (MAP) image and a cross-sectional image of skin pores in the facial area. Subsequently, we detected the sebum in skin pores using the detection algorithm of the ImageJ software to quantitatively determine the size of randomly selected pores in the proposed MAP images. Additionally, the pore size was analyzed by acquiring images before and after facial sebum extraction. According to our research, facial sebum can be morphologically described using the OCT system. Since OCT imaging enables specific analysis of skin parameters, including pores and sebum, skin analysis employing OCT could be an effective method for further research.

Metabolically distinct roles of NAD synthetase and NAD kinase define the essentiality of NAD and NADP in Mycobacterium tuberculosis.

Nicotinamide adenine dinucleotide (NAD) and its phosphorylated derivative (NADP) are essential cofactors that participate in hundreds of biochemical reactions and have emerged as therapeutic targets in cancer, metabolic disorders, neurodegenerative diseases, and infections, including tuberculosis. The biological basis for the essentiality of NAD(P) in most settings, however, remains experimentally unexplained. Here, we report that inactivation of the terminal enzyme of NAD synthesis, NAD synthetase (NadE), elicits markedly different metabolic and microbiologic effects than those of the terminal enzyme of NADP biosynthesis, NAD kinase (PpnK), in Mycobacterium tuberculosis (Mtb). Inactivation of NadE led to parallel reductions of both NAD and NADP pools and Mtb viability, while inactivation of PpnK selectively depleted NADP pools but only arrested growth. Inactivation of each enzyme was accompanied by metabolic changes that were specific for the affected enzyme and associated microbiological phenotype. Bacteriostatic levels of NAD depletion caused a compensatory remodeling of NAD-dependent metabolic pathways in the absence of an impact on NADH/NAD ratios, while bactericidal levels of NAD depletion resulted in a disruption of NADH/NAD ratios and inhibition of oxygen respiration. These findings reveal a previously unrecognized physiologic specificity associated with the essentiality of two evolutionarily ubiquitous cofactors. IMPORTANCE The current course for cure of Mycobacterium tuberculosis (Mtb)-the etiologic agent of tuberculosis (TB)-infections is lengthy and requires multiple antibiotics. The development of shorter, simpler treatment regimens is, therefore, critical to the goal of eradicating TB. NadE, an enzyme required for the synthesis of the ubiquitous cofactor NAD, is essential for survival of Mtb and regarded as a promising drug target. However, the basis of this essentiality was not clear due to its role in the synthesis of both NAD and NADP. Here, we resolve this ambiguity through a combination of gene silencing and metabolomics. We specifically show that NADP deficiency is bacteriostatic, while NAD deficiency is bactericidal due to its role in Mtb's respiratory capacity. These results argue for a prioritization of NAD biosynthesis inhibitors in anti-TB drug development.

Cook with Different Pots, but Similar Taste? Comparison of Phase Angle Using Bioelectrical Impedance Analysis According to Device Type and Examination Posture.

Bioelectrical impedance analysis (BIA) is gaining popularity as a tool for body composition assessment. Although BIA has been studied and validated in different populations, age groups, and clinical settings, including critically ill patients, there are concerns about BIA reproducibility and reliability for different device types and postures. This study aimed to evaluate the reliability of BIA using different devices, postures, and lead types. Cross-sectional observational data were collected from 74 healthy volunteers (32 women, 42 men). We used two types of devices, three types of postures (standing, sitting, and lying), and two lead types (clamp lead and adhesive lead) to measure the whole-body phase angle (phA) at a single frequency of 50 kHz. The measurements were validated using the intraclass correlation coefficient (ICC) and Bland-Altman plot analysis. All phA measurements recorded using the two types of devices, three different postures, and two types of leads were equivalent (mean ICC = 0.9932, 95% confidence interval (CI) 0.9905-0.0053, p < 0.001). The average mean difference in phA was 0.31 (95% CI 0.16-0.46). The largest phA value was measured using BWA with an adhesive-type lead in the supine position. There were no differences between the standing and sitting positions. We compared the consistency and reliability of phA using two devices, two lead types, and three postures. Seven different phA were interchangeable in healthy volunteers.

Genotypes and phenotypes of DNM1 encephalopathy.

BACKGROUND: Variants in the dynamin-1 (DNM1) gene typically cause synaptopathy, leading to developmental and epileptic encephalopathy (DEE). We aimed to determine the genotypic and phenotypic spectrum of DNM1 encephalopathy beyond DEE. METHODS: Electroclinical phenotyping and genotyping of patients with a DNM1 variant were conducted for patients undergoing next-generation sequencing at our centre, followed by a systematic review. RESULTS: Six patients with heterozygous DNM1 variants were identified in our cohort. Three had a typical DEE phenotype characterised by epileptic spasms, tonic seizures and severe-to-profound intellectual disability with pathogenic variants located in the GTPase or middle domain. The other three patients had atypical phenotypes of milder cognitive impairment and focal epilepsy. Genotypically, two patients with atypical phenotypes had variants located in the GTPase domain, while the third patient had a novel variant (p.M648R) in the linker region between pleckstrin homology and GTPase effector domains. The third patient with an atypical phenotype showed normal development until he developed febrile status epilepticus. Our systematic review on 55 reported cases revealed that those with GTPase or middle domain variants had more severe intellectual disability (p<0.001) and lower functional levels of ambulation (p=0.001) or speech and language (p<0.001) than the rest. CONCLUSION: DNM1-related phenotypes encompass a wide spectrum of epilepsy and neurodevelopmental disorders, with specific variants underlying different phenotypes.

Lymphocyte-activating gene 3 expression in tumor cells predicts immune checkpoint inhibitor response in triple negative breast cancer.

Introduction: Immune checkpoint inhibitor (ICI) is one of the standard treatment strategies in triple negative breast cancer (TNBC). However, the benefit of ICI with chemotherapy is limited in metastatic TNBC. In this study, we evaluated the effect of PD-L1 and LAG-3 expression on tissue microenvironment of mTNBC treated with ICI. Methods: We reviewed representative formalin-fixed paraffin embedded specimens from metastatic or archival tumor tissues of TNBCs who treated with PD-1/PD-L1 inhibitors in metastatic setting. We used the Opal multiplex Detection kit with six antibodies (anti-PD-L1, anti-LAG-3, anti-CD68, anti-panCK, anti-CD8, anti-CD107a/LAMP antibody). Results: We evaluated the association between LAG-3+cells and survival outcome regarding CK expression. Stromal LAG-3+/CK+ and LAG-3+/CK- cells were not associated with ICI-progression free survival(PFS) (P=0.16). However, LAG-3+ cell distributions in the tumor area impacted on ICI-PFS. A high density of LAG-3+CK+ cells was associated with shorter ICI-PFS compared with low densities of both LAG-3+CK+ and LAG-3+CK- cells (1.9 vs. 3.5 months). In addition, a high density of LAG-3+CK- cells had a relatively longer ICI-PFS compared with other groups (P=0.01). In terms of total area, the pattern of densities of LAG-3+CK+ cells and LAG-3+CK- cells were similar to those in the tumor area In addition, ICI-PFS of LAG-3+CK- and LAG-3+CK+ cell densities in the total area was equal to that in the tumor area. Discussion: In conclusion, our findings revealed tumor-intrinsic LAG-3 expression was the resistance mechanism toward PD-1/PD-L1 inhibitors in mTNBCs. Multivariate analysis also suggested that LAG-3 expression in tumor cells was an independent predictive biomarker.

Integrating simultaneous motor imagery and spatial attention for EEG-BCI control.

OBJECTIVE: EEG-based brain-computer interfaces (BCI) are non-invasive approaches for replacing or restoring motor functions in impaired patients, and direct brain-to-device communication in the general population. Motor imagery (MI) is one of the most used BCI paradigms, but its performance varies across individuals and certain users require substantial training to develop control. In this study, we propose to integrate a MI paradigm simultaneously with a recently proposed Overt Spatial Attention (OSA) paradigm, to accomplish BCI control. METHODS: We evaluated a cohort of 25 human subjects' ability to control a virtual cursor in one- and two-dimensions over 5 BCI sessions. The subjects used 5 different BCI paradigms: MI alone, OSA alone, MI and OSA simultaneously towards the same target (MI+OSA), and MI for one axis while OSA controls the other (MI/OSA and OSA/MI). RESULTS: Our results show that MI+OSA reached the highest average online performance in 2D tasks at 49% Percent Valid Correct (PVC), statistically outperforms MI alone (42%), and was higher, but not statistically significant, than OSA alone (45%). MI+OSA had a similar performance to each subject's best individual method between MI alone and OSA alone (50%) and 9 subjects reached their highest average BCI performance using MI+OSA. CONCLUSION: Integrating MI and OSA leads to improved performance over MI alone at the group level and is the best BCI paradigm option for some subjects. SIGNIFICANCE: This work proposes a new BCI control paradigm that integrates two existing paradigms and demonstrates its value by showing that it can improve users' BCI performance.

Development of a deviated focusing-based optical coherence microscope with a variable depth of focus for high-resolution imaging.

The aim of this study was to develop an optically deviated focusing-based variable depth-of-focus (DOF) oriented optical coherence microscopy (OCM) system to improve the DOF in high-resolution and precise focused imaging. In this study, an approach of varying beam diameter using deviated focusing was employed in the sample arm to enhance the DOF and to confirm precise focusing in OCM imaging. The optically deviated focusing technique was used to vary the focal point and DOF by altering the sample arm beam. The efficacy of the variable DOF imaging approach utilizing an optimized sample arm was confirmed by tissue-level imaging, where OCM images with varying DOF were obtained using deviated focusing. Experimentally confirmed lateral resolution of 2.19 µm was sufficient for the precise non-invasive visualization of abnormalities of fruit specimens. Thus, the proposed variable DOF-OCM system can be an alternative for precisely focused, high-resolution, and variable DOF imaging by improving the DOF in minimum lateral resolution variation.

Health behaviors and the risk of COVID-19 incidence: A Bayesian hierarchical spatial analysis.

OBJECTIVES: Effective infection control measures, based on a sound understanding of geographical community-specific health behavioral characteristics, should be implemented from the early stage of disease transmission. However, few studies have explored health behaviors as a possible contributor to COVID-19 infection in the spatial context. We investigated health behaviors as potential factors of COVID-19 incidence in the early phase of transmission in the spatial context. METHODS: We extracted COVID-19 cumulative case data as of February 25, 2021-one day prior to nationwide COVID-19 vaccination commencement-regarding health behaviors and covariates, including health condition and socio-economic factors, at the municipal level from publicly available datasets. The spatial autocorrelation of incidence was analyzed using Global Moran's I statistics. The associations between health behaviors and COVID-19 incidence were examined using Besag-York-Mollie models to deal with spatial autocorrelation of residuals. RESULTS: The COVID-19 incidence had positive spatial autocorrelation across South Korea (I = 0.584, p = 0.001). The results suggest that individuals vaccinated against influenza in the preceding year had a negative association with COVID-19 incidence (relative risk=0.913, 95 % Credible Interval=0.838-0.997), even after adjusting for covariates. CONCLUSIONS: Our ecological study suggests an association between COVID-19 infection and health behaviors, especially influenza vaccination, in the early stage of COVID-19 transmission at the municipal level.

Dental diagnosis for inlay restoration using an intraoral optical coherence tomography system: A case report.

PATIENTS: The patient was a 32-year-old man who underwent amalgam restoration of the mandibular right second molar. An amalgam restoration fracture was diagnosed by intraoral optical coherence tomography (OCT), and pulp exposure was examined during cavity preparation. Subsequently, a definitive ceramic restoration was fabricated, and the marginal fit in the oral cavity was evaluated using the OCT system. DISCUSSION: The existing OCT system cannot acquire images inside the oral cavity because of the large probe size. However, the proposed intraoral OCT system can access the prostheses in the mandibular right second molar. Therefore, dental diagnosis for restoration treatment with dental prosthesis fracture, marginal gap, and pulp exposure after tooth preparation is possible using the proposed intraoral OCT system. CONCLUSIONS: The use of the intraoral OCT system improved dental diagnosis by allowing the dentist to confirm quantitative values through cross-sectional images, rather than that by determining a treatment plan after visual dental diagnosis.

Three-dimensional reconstructing undersampled photoacoustic microscopy images using deep learning.

Spatial sampling density and data size are important determinants of the imaging speed of photoacoustic microscopy (PAM). Therefore, undersampling methods that reduce the number of scanning points are typically adopted to enhance the imaging speed of PAM by increasing the scanning step size. Since undersampling methods sacrifice spatial sampling density, by considering the number of data points, data size, and the characteristics of PAM that provides three-dimensional (3D) volume data, in this study, we newly reported deep learning-based fully reconstructing the undersampled 3D PAM data. The results of quantitative analyses demonstrate that the proposed method exhibits robustness and outperforms interpolation-based reconstruction methods at various undersampling ratios, enhancing the PAM system performance with 80-times faster-imaging speed and 800-times lower data size. The proposed method is demonstrated to be the closest model that can be used under experimental conditions, effectively shortening the imaging time with significantly reduced data size for processing.

Optical Coherence Tomography as a Non-Invasive Tool for Plant Material Characterization in Agriculture: A Review.

Characterizing plant material is crucial in terms of early disease detection, pest control, physiological assessments, and growth monitoring, which are essential parameters to increase production in agriculture and prevent unnecessary economic losses. The conventional methods employed to assess the aforementioned parameters have several limitations, such as invasive inspection, complexity, high time consumption, and costly features. In recent years, optical coherence tomography (OCT), which is an ultra-high resolution, non-invasive, and real-time unique image-based approach has been widely utilized as a significant and potential tool for assessing plant materials in numerous aspects. The obtained OCT cross-sections and volumetrics, as well as the amplitude signals of plant materials, have the capability to reveal vital information in both axial and lateral directions owing to the high resolution of the imaging system. This review discusses recent technological trends and advanced applications of OCT, which have been potentially adapted for numerous agricultural applications, such as non-invasive disease screening, optical signals-based growth speed detection, the structural analysis of plant materials, and microbiological discoveries. Therefore, this review offers a comprehensive exploration of recent advanced OCT technological approaches for agricultural applications, which provides insights into their potential to incorporate OCT technology into numerous industries.

On-field optical imaging data for the pre-identification and estimation of leaf deformities.

Visually nonidentifiable pathological symptoms at an early stage are a major limitation in agricultural plantations. Thickness reduction in palisade parenchyma (PP) and spongy parenchyma (SP) layers is one of the most common symptoms that occur at the early stage of leaf diseases, particularly in apple and persimmon. To visualize variations in PP and SP thickness, we used optical coherence tomography (OCT)-based imaging and analyzed the acquired datasets to determine the threshold parameters for pre-identifying and estimating persimmon and apple leaf abnormalities using an intensity-based depth profiling algorithm. The algorithm identified morphological differences between healthy, apparently-healthy, and infected leaves by applying a threshold in depth profiling to classify them. The qualitative and quantitative results revealed changes and abnormalities in leaf morphology in addition to disease incubation in both apple and persimmon leaves. These can be used to examine how initial symptoms are influenced by disease growth. Thus, these datasets confirm the significance of OCT in identifying disease symptoms nondestructively and providing a benchmark dataset to the agriculture community for future reference.

Association between Climate Factors and Dengue Fever in Asuncion, Paraguay: A Generalized Additive Model.

Dengue fever has been endemic in Paraguay since 2009 and is a major cause of public-health-management-related burdens. However, Paraguay still lacks information on the association between climate factors and dengue fever. We aimed to investigate the association between climatic factors and dengue fever in Asuncion. Cumulative dengue cases from January 2014 to December 2020 were extracted weekly, and new cases and incidence rates of dengue fever were calculated. Climate factor data were aggregated weekly, associations between dengue cases and climate factors were analyzed, and variables were selected to construct our model. A generalized additive model was used, and the best model was selected based on Akaike information criteria. Piecewise regression analyses were performed for non-linear climate factors. Wind and relative humidity were negatively associated with dengue cases, and minimum temperature was positively associated with dengue cases when the temperature was less than 21.3 °C and negatively associated with dengue when greater than 21.3 °C. Additional studies on dengue fever in Asuncion and other cities are needed to better understand dengue fever.

Accumulated Vitrified Embryos Could Be a Method for Increasing Pregnancy Rates in Patients with Poor Ovarian Response.

We aimed to assess the efficacy of accumulated embryo transfer (ACC-ET) through several controlled ovarian hyperstimulation (COS) cycles to increase the rates of pregnancy in patients with poor ovarian response (POR). We retrospectively reviewed the medical records of 588 patients with POR under 43-years old who underwent embryo transfer from January 2010 to December 2015. We compared the pregnancy rate (PR), clinical pregnancy rate (CPR), and live birth rate (LBR) between ACC-ET (frozen-thawed: 47; fresh + frozen-thawed: 24) group (n = 71) and fresh ET groups (n = 517). Characteristics of ACC-ET patients were similar to those of fresh ET groups (Age: 38.1 ± 3.5 vs. 38.2 ± 3.7, p = 0.88; Anti Müllerian Hormone (AMH; ng/mL): 0.5 ± 0.4 vs. 0.6 ± 0.6, p = 0.38; follicle stimulating hormone (FSH: mIU/mL): 11.9 ± 8.0 vs. 10.8 ± 9.0, p = 0.35). The total number of transferred embryos (3.1 ± 0.9 vs. 1.5 ± 0.7, p = 0.00), PR (29.6% (21/71) vs. 18.8% (97/517), p = 0.040), and CPR (23.5% (16/68) vs. 14.0% (71/508) p = 0.047) were significantly higher in the ACC-ET group than in the fresh ET group. In addition, PR, CPR, and LBR increased with the number of ET in the fresh ET group. However, there were no significant differences observed in LBR between ACC-ET and fresh ET groups (14.9% (10/67) vs. 9.8% (50/508), p = 0.203). From our knowledge, there is no clinical evidence reported to prove that transfer of multiple embryos of adequate quality obtained through multiple cycles can compensate for the limited number of retrieved oocytes from POR patients. We concluded that ACC-ET from several COS cycles could be an alternative method to increase PR and CPR in <43-year-old patients with POR.

Effects of Elevated Progesterone Levels on the Day of hCG on the Quality of Oocyte and Embryo.

This study is designed to investigate the effects of increased progesterone (P4) levels on the quality of retrieved oocytes and embryos during IVF. This retrospective analysis included 982 all-freezing in vitro fertilization (IVF) cycles (conducted between November 2019 and June 2020 at CHA Fertility Center Bundang, South Korea) in which serum P4 levels were measured on the day of human chorionic gonadotropin (hCG) administration. Our study revealed that the serum P4 levels on the day of hCG administration are strongly associated with the rates of oocyte maturation, displaying a positive correlation in patients with serum P4 < 2.25 ng/mL (p = 0.025). Moreover, patients with serum P4 < 1.25 ng/mL showed relatively low fertilization rates (p = 0.037), and the rates of good embryo retrieval were significantly increased with the serum P4 level < 1.5 ng/mL (p = 0.001). Interestingly, serum P4 level on the day of hCG administration affects the rate of good-quality embryo development, especially at the cleavage stage, and is associated with the status of ovarian responses. Our current study suggests that serum P4 level on the day of hCG administration negatively affects the rates of oocyte maturation, fertilization, and the development of good embryos.

Target ischemic stroke model creation method using photoacoustic microscopy with simultaneous vessel monitoring and dynamic photothrombosis induction.

The ischemic stroke animal model evaluates the efficacy of reperfusion and neuroprotective strategies for ischemic injuries. Various conventional methods have been reported to induce the ischemic models; however, controlling specific neurological deficits, mortality rates, and the extent of the infarction is difficult as the size of the affected region is not precisely controlled. In this paper, we report a single laser-based localized target ischemic stroke model development method by simultaneous vessel monitoring and photothrombosis induction using photoacoustic microscopy (PAM), which has minimized the infarct size at precise location with high reproducibility. The proposed method has significantly reduced the infarcted region by illuminating the precise localization. The reproducibility and validity of suggested method have been demonstrated through repeated experiments and histological analyses. These results demonstrate that our method can provide the ischemic stroke model closest to the clinical pathology for brain ischemia research from inducement, occurrence mechanisms to the recovery process.