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MACS-W:A modified optical clearing agent for imaging 3D cell cultures

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Journal of Innovative Optical Health sciences 2024年第2期17卷 24-34页

作者： Xiang Zhong Chao Gao Hui Li Yuening He Peng Fei Zaozao Chen Zhongze Gu Dan Zhu Tingting Yu britton chance center for biomedical photonics-moe key laboratory for biomedical photonics huazhong university of science and technology wuhan Hubei 430074P.R.China wuhan National laboratory for Optoelectronics-Advanced biomedical Imaging Facility huazhong university of science and technology wuhan Hubei 430074P.R.China School of Optical and Electronic Infrmation huazhong university of science and technology wuhan Hubei 430074P.R.China State key laboratory of Bioelectronics School of Biological science and Medical Engineering Southeast university NanjingJiangsu 210096P.R.China Institute of Biomaterials and Medical Devices Southeast university Suzhou Jiangsu 215163P.R.China

Three-dimensional(3D)cell cultures have contributed to a variety of biological research fields by filling the gap between monolayers and animal models.The modern optical sectioning microscopic methods make it possible to probe the complexity of 3D cell cultures but are limited by the inherent opaqueness.While tissue optical clearing methods have emerged as powerful tools for investigating whole-mount tissues in 3D,they often have limitations,such as being too harsh for fragile 3D cell cultures,requiring complex handling protocols,or inducing tissue deformation with shrinkage or expansion.To address this issue,we proposed a modified optical clearing method for 3D cell cultures,called MACS-W,which is simple,highly efficient,and morphology-preserving.In our evaluation of MACS-W,we found that it exhibits excellent clearing capability in just 10 min,with minimal deformation,and helps drug evaluation on tumor spheroids.In summary,MACS-W is a fast,minimally-deformative and fluorescence compatible clearing method that has the potential to be widely used in the studies of 3D cell cultures.

关键词： Tissue optical clearing 3D cell cultures imaging

Tissue optical clearing enhances efficacy of vascular targeted photodynamic therapy of mouse dorsal skin

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Journal of Innovative Optical Health sciences 2024年第2期17卷 67-78页

作者： Ying Liu Qiushi Wang Yidi Liu Ying Wang Haixia Qiu Dan Zhu Ying Gu Defu Chen School of Optics and photonics Beijing Institute of TechnologyBeijing 100081P.R.China School of Medical technology Beijing Institute of TechnologyBeijing 100081P.R.China Department of Laser Medicine First Medical Center of PLA General HospitalBeijing 100853P.R.China britton chance center for biomedical photonics-moe key laboratory for biomedical photonics A duanced Biomedical Imaging FacilityWuhan National Laboratory for OptoelectronicsHuazhong University of Science and TechnologyWuhan 430074HubeiP.R.China Precision Laser Medical Diagnosis and Treatment Innovation Unit Chinese A cademy of Medical SciencesBeijing 100000P.R.China

Vascular-targeted photodynamic therapy(V-PDT)is an effective treatment for port wine stains(PWS).However,repeated treatment is usually needed to achieve optimal treatment outcomes,possibly due to the limited treatment light penetration depth in the PWS lesion.The optical clearing technique can increase light penetration in depth by reducing light scattering.This study aimed to investigate the V-PDT in combination with an optical clearing agent(OCA)for the therapeutic enhancement of V-PDT in the rodent skinfold window chamber model.Vascular responses were closely monitored with laser speckle contrast imaging(LSCI),optical coherence tomography angiography,and stereo microscope before,during,and after the treatment.We further quantitatively demonstrated the effects of V-PDT in combination with OCA on the blood flow and blood vessel size of skin microvasculature.The combination of OCA and V-PDT resulted in significant vascular damage,including vasoconstriction and the reduction of blood flow.Our results indicate the promising potential of OCA for enhancing V-PDT for treating vascular-related diseases,including PWS.

关键词： Vascular-targeted photodynamic therapy(V-PDT) optical clearing agent(OCA) treatment efficacy enhancement skin-fold window chamber port wine stains

Acceptor Engineering Produces Ultrafast Nonradiative Decay in NIR-Ⅱ Aza-BODIPY Nanoparticles for Efficient Osteosarcoma Photothermal Therapy via Concurrent Apoptosis and Pyroptosis

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Research 2024年第1期2024卷 625-635页

作者： Zhenxiong Shi Hua Bai Jiaxing Wu Xiaofei Miao Jia Gao Xianning Xu Yi Liu Jiamin Jiang Jiaqi Yang Jiaxin Zhang Tao Shao Bo Peng Huili Ma Dan Zhu Guojing Chen Wenbo Hu Lin Li Wei Huang Frontiers science center for Flexible Electronics Xi’an Institute of Flexible Electronics(IFE)and Xi’an Institute of Biomedical Materials&EngineeringNorthwestern Polytechnical UniversityXi’an 710072China key laboratory for Organic Electronics and Information Displays&Institute of Advanced Materials(IAM) Nanjing University of Posts&TelecommunicationsNanjing 210023China key laboratory of Flexible Electronics(KLOFE)and IAM Nanjing Tech UniversityNanjing 211800China britton chance center for biomedical photonics-moe key laboratory for biomedical photonics Wuhan National Laboratory for Optoelectronics-Advanced Biomedical Imaging FacilityHuazhong University of Science and TechnologyWuhan 430074China Department of Orthopedics Xijing HospitalThe Fourth Military Medical UniversityXi’an 710032China The Institute of Flexible Electronics(IFE Future Technologies)Xiamen UniversityXiamen 361005China

Small-molecule photothermal agents(PTAs)with intense second near-infrared(NIR-Ⅱ,1,000 to 1,700 nm)absorption and high photothermal conversion efficiencies(PCEs)are promising candidates for treating deep-seated tumors such as osteosarcoma.To date,the development of small-molecule NIR-Ⅱ PTAs has largely relied on fabricating donor–acceptor–donor(D–A–D/D′)structures and limited success has been achieved.Herein,through acceptor engineering,a donor–acceptor–acceptor(D–A–A′)-structured NIR-Ⅱ aza-boron-dipyrromethene(aza-BODIPY)PTA(SW8)was readily developed for the 1,064-nm laser-mediated phototheranostic treatment of osteosarcoma.Changing the donor groups to acceptor groups produced remarkable red-shifts of absorption maximums from first near-infrared(NIR-Ⅰ)regions(~808 nm)to NIR-Ⅱ ones(~1,064 nm)for aza-BODIPYs(SW1 to SW8).Furthermore,SW8 self-assembled into nanoparticles(SW8@NPs)with intense NIR-Ⅱ absorption and an ultrahigh PCE(75%,1,064 nm).This ultrahigh PCE primarily originated from an additional nonradiative decay pathway,which showed a 100-fold enhanced decay rate compared to that shown by conventional pathways such as internal conversion and vibrational relaxation.Eventually,SW8@NPs performed highly efficient 1,064-nm laser-mediated NIR-Ⅱ photothermal therapy of osteosarcoma via concurrent apoptosis and pyroptosis.This work not only illustrates a remote approach for treating deep-seated tumors with high spatiotemporal control but also provides a new strategy for building high-performance small-molecule NIR-Ⅱ PTAs.

关键词： donor BODIPY intense

Organic fluorescent probes for live-cell super-resolution imaging

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Frontiers of Optoelectronics 2023年第4期16卷 25-36页

作者： Xinxin Duan Meng Zhang Yu‑Hui Zhang britton chance center for biomedical photonics MoE Key Laboratory for Biomedical PhotonicsAdvanced Biomedical Imaging Facility-Wuhan National Laboratory for OptoelectronicsHuazhong University of Science and TechnologyWuhan 430074China

The development of super-resolution technology has made it possible to investigate the ultrastructure of intracellular organelles by fuorescence microscopy,which has greatly facilitated the development of life sciences and biomedicine.To realize super-resolution imaging of living cells,both advanced imaging systems and excellent fuorescent probes are required.Traditional fuorescent probes have good availability,but that is not the case for probes for live-cell super-resolution imaging.In this review,we frst introduce the principles of various super-resolution technologies and their probe requirements,then summarize the existing designs and delivery strategies of super-resolution probes for live-cell imaging,and fnally provide a brief conclusion and overview of the future.

关键词： Super-resolution imaging Organic fluorescent dyes-Live-cell imaging Cell-impermeable organic probes

Photostimulation of lymphatic clearance ofβ‑amyloid from mouse brain:a new strategy for the therapy of Alzheimer’s disease

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Frontiers of Optoelectronics 2023年第4期16卷 179-190页

作者： Dongyu Li Hao Lin Silin Sun Shaojun Liu Zhang Liu Yuening He Jingtan Zhu Jianyi Xu Oxana Semyachkina‑Glushkovskaya Tingting Yu Dan Zhu britton chance center for biomedical photonics-moe key laboratory for biomedical photonics Wuhan National Laboratory for Optoelectronics-Advanced Biomedical Imaging FacilityHuazhong University of Science and TechnologyWuhan 430074China School of Optical Electronic Information Huazhong University of Science and TechnologyWuhan 430074China Department of Biology Saratov State UniversitySaratov 410012Russia

Alzheimer’s disease(AD)is an age-related neurodegenerative disorder that poses a signifcant burden on socio-economic and healthcare systems worldwide.However,the currently available therapy of AD is limited,and new strategies are needed to enhance the clearance ofβ-amyloid(Aβ)protein and improve cognitive function.Photobiomodulation(PBM)is a noninvasive and efective therapeutic method that has shown promise in treating various brain diseases.Here,we demonstrate that 1267-nm PBM signifcantly alleviates cognitive decline in the 5xFAD mouse model of AD and is safe as it does not induce a signifcant increase in cortical temperature.Moreover,with the combination of 3D tissue optical clearing imaging and automatic brain region segmentation,we show that PBM-mediated reductions of Aβplaques in diferent subregions of prefrontal cortex and the hippocampus are diferent.The PBM-induced lymphatic clearance of Aβfrom the brain is associated with improvement of memory and cognitive functions in 5xFAD mice.Our results suggest that the modulation of meningeal lymphatic vessels(MLVs)should play an important role in promoting Aβclearance.Collectively,this pilot study demonstrates that PBM can safely accelerate lymphatic clearance of Aβfrom the brain of 5xFAD mice,promoting improvement of neurocognitive status of AD animals suggesting that PBM can be an efective and bedside therapy for AD.

关键词： Photostimulation-Meningeal lymphatic vessels Amyloid-βclearance Alzheimer's disease

Constructing the rodent stereotaxic brain atlas:a survey

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science China(Life sciences) 2022年第1期65卷 93-106页

作者： Zhao Feng Anan Li Hui Gong Qingming Luo britton chance center for biomedical photonics Wuhan National Laboratory for OptoelectronicsMOE Key Laboratory for Biomedical PhotonicsHuazhong University of Science and TechnologyWuhan 430074China HUST-Suzhou Institute for Brainsmatics Suzhou 215123China School of biomedical Engineering Hainan UniversityHaikou 570228China

The stereotaxic brain atlas is a fundamental reference tool commonly used in the field of neuroscience.Here we provide a brief history of brain atlas development and clarify three key conceptual elements of stereotaxic brain atlasing:brain image,atlas,and stereotaxis.We also refine four technical indices for evaluating the construction of atlases:the quality of staining and labeling,the granularity of delineation,spatial resolution,and the precision of spatial location and orientation.Additionally,we discuss state-of-the-art technologies and their trends in the fields of image acquisition,stereotaxic coordinate construction,image processing,anatomical structure recognition,and publishing:the procedures of brain atlas illustration.We believe that the use of single-cell resolution and micron-level location precision will become a future trend in the study of the stereotaxic brain atlas,which will greatly benefit the development of neuroscience.

关键词： brain atlas stereotaxic cytoarchitecture brainsmatics

C1M2:a universal algorithm for 3D instance segmentation,annotation,and quantification of irregular cells

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science China(Life sciences) 2023年第10期66卷 2415-2428页

作者： Hao Zheng Songlin Huang Jing Zhang Ren Zhang Jialu Wang Jing Yuan Anan Li Xin Yang Zhihong Zhang britton chance center and moe key laboratory for biomedical photonics Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and TechnologyWuhan 430074China key laboratory of biomedical Engineering of Hainan Province School of Biomedical EngineeringHainan UniversityHaikou 570228China School of Electronic Information and Communications Huazhong University of Science and TechnologyWuhan 430074China

Cell instance segmentation is a fundamental task for many biological applications,especially for packed cells in three-dimensional(3D)microscope images that can fully display cellular morphology.Image processing algorithms based on neural networks and feature engineering have enabled great progress in two-dimensional(2D)instance segmentation.However,current methods cannot achieve high segmentation accuracy for irregular cells in 3D images.In this study,we introduce a universal,morphology-based 3D instance segmentation algorithm called Crop Once Merge Twice(C1M2),which can segment cells from a wide range of image types and does not require nucleus images.C1M2 can be extended to quantify the fluorescence intensity of fluorescent proteins and antibodies and automatically annotate their expression levels in individual cells.Our results suggest that C1M2 can serve as a tissue cytometry for 3D histopathological assays by quantifying fluorescence intensity with spatial localization and morphological information.

关键词： 3D instance segmentation irregular cells fluorescence images neural networks fluorescence intensity tissue cytometry

Tissue optical clearing for neural regeneration research

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Neural Regeneration Research 2023年第9期18卷 1940-1941页

作者： Tingting Yu Jianyi Xu Dan Zhu britton chance center for biomedical photonics-moe key laboratory for biomedical photonics wuhan National laboratory for Optoelectronics-Advanced biomedical Imaging Facility Huazhong University of Science and TechnologyWuhanHubei ProvinceChina Optics Valley laboratory WuhanHubei ProvinceChina

Nerve injury,whether traumatic or degenerative,disrupts the transmission of information in the nervous syste m,leading to dysfunction.It is widely known that neural regeneration is vital to the restoration of function after nerve injury.Still,outcomes are often limited by the misguidance of axonal regeneration and complex pathological changes in neurons and glia,as well as the longterm denervation of target organs.

关键词： organs neural traumatic

Whole-Brain Direct Inputs to and Axonal Projections from Excitatory and Inhibitory Neurons in the Mouse Primary Auditory Area

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Neuroscience Bulletin 2022年第6期38卷 576-590页

作者： Mengting Zhao Miao Ren Tao Jiang Xueyan Jia Xiaojun Wang Anan Li Xiangning Li Qingming Luo Hui Gong Briton chance center for biomedical photonics Wuhan National Laboratory for OptoelectronicsMoE Key Laboratory for Biomedical PhotonicsHuazhong University of Science and TechnologyWuhan 430074China key laboratory of biomedical Engineering of Hainan Province School of Biomedical EngineeringHainan UniversityHaikou 570228China HUST-Suzhou Institute for Brainsmatics JITRISuzhou 215123China

Neurons in the primary auditory area(AUDp)innervate multiple brain regions with long-range projections while receiving informative inputs for diverse functions.However,the brain-wide connections of these neurons have not been comprehensively investigated.Here,we simultaneously applied virus-based anterograde and retrograde tracing,labeled the connections of excitatory and inhibitory neurons in the mouse AUDp,and acquired whole-brain information using a dual-channel fuorescence micro-optical sectioning tomography system.Quantified results showed that the two types of neurons received inputs with similar patterns but sent heterogeneous projections to downstream regions.In the isocortex,functionally different areas consistently sent feedback-dominated projections to these neurons,with concomitant laterallydominated projections from the sensory and limbic cortices to inhibitory neurons.In subcortical regions,the dorsal and medial parts of the non-lemniscal auditory thalamus(AT)were reciprocally connected to the AUDp,while the ventral part contained the most fibers of passage from the excitatory neurons and barely sent projections back,indicating the regional heterogeneity of the AUDp-AT circuit.Our results reveal details of the whole-brain network and provide new insights for further physiological and functional studies of the AUDp.

关键词： Primary auditory area Excitatory Inhibitory Direct inputs Axonal projections