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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

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

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

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

前背侧终纹床核中与焦虑相关的细胞类型特异性神经环路图谱的研究

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Science Bulletin 2020年第14期65卷 1203-1216,M0004页

作者：王鑫鑫张勇升王旭代家起化瑞芳曾绍群李浩洪 Britton Chance Center for Biomedical Photonics Wuhan National Laboratory for OptoelectronicsHuazhong University of Science and TechnologyWuhan 430074China Ministry of Education Key Laboratory for biomedical photonics Collaborative Innovation Center for Biomedical EngineeringSchool of Engineering SciencesHuazhong University of Science and TechnologyWuhan 430074China

焦虑是生物个体面临外界环境潜在威胁时的本能反应.既往研究表明终纹床核(the bed nucleus of the stria terminalis, BNST)在调节焦虑过程中起着重要作用,但其中与焦虑行为相关的神经环路构筑和功能尚未可知.本研究在前背侧终纹床核(an... 详细信息

焦虑是生物个体面临外界环境潜在威胁时的本能反应.既往研究表明终纹床核(the bed nucleus of the stria terminalis, BNST)在调节焦虑过程中起着重要作用,但其中与焦虑行为相关的神经环路构筑和功能尚未可知.本研究在前背侧终纹床核(anterior-dorsal BNST, ad BNST)中发现两类独立分布的细胞群体,分别特异性表达促肾上腺皮质激素释放激素(corticotropin-releasing hormone, CRH)和蛋白激酶C-δ(protein kinase C-δ, PKC-δ)分子.利用神经环路示踪、全细胞膜片钳、在体神经元钙离子成像以及化学遗传学手段,本文阐明了ad BNST的CRH与PKC-δ阳性神经元的局部和长距离的神经环路连接特点及其对小鼠焦虑行为产生的不同调控作用.研究结果表明, CRH与PKC-δ神经元接收相似的上游脑区神经投射,并在部分下游脑区的投射密度中表现出显著性差异.该结果提示,二者可能参与到不同的焦虑行为调控中.进一步的功能研究结果发现,特异性地操纵CRH和PKC-δ阳性神经元能够分别产生致焦虑和抗焦虑作用.本研究揭示了ad BNST脑区CRH与PKC-δ神经元的功能异质性,为后续调控焦虑的神经环路机制的研究提供了坚实的基础.

关键词：促肾上腺皮质激素释放激素神经环路全细胞膜片钳终纹床核阳性神经元本能反应生物个体蛋白激酶C

A brief introduction to biophotonic techniques and methods

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Science China(Life Sciences) 2020年第12期63卷 1771-1775页

作者： Qingming Luo School of biomedical Engineering Hainan UniversityHaikou 570228China Britton Chance Center for Biomedical Photonics MoE Key Laboratory for Biomedical PhotonicsWuhan National Laboratory for Optoelectronics-Huazhong University of Science and TechnologyWuhan 430074China

Biophotonics is not a standard word listed in a classical dictionary. The meaning of "biophotonics" is essentially the use of photonic technologies to detect and perhaps to control a biological phenomenon or event, at different levels ranging from molecules, cells, tissue, organisms, to intact animals and even human bodies(Yeh and Krishnan, 2018). The field of biophotonics is not well defined because it is a multidisciplinary research field at the intersection of physics,biology, chemistry, medicine and engineering(Tsia, 2016;Vo-Dinh, 2014;Wang and Tuchin, 2016). As alternatives to biophotonics, the term "biomedical photonics" and "biomedical optics" have also been popularly used, with no exact distinction among them(Vo-Dinh, 2014).

关键词： listed optics exact

Tissue optical clearing imaging for structural changes of neuromuscular junctions after mice ischemic stroke[Invited]

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Chinese Optics Letters 2023年第12期21卷 54-60页

作者：徐健壹戴奕宣昂姚应涛刘少军俞婷婷朱䒟 Britton Chance Center for Biomedical Photonics MoE Key Laboratory for Biomedical PhotonicsWuhan National Laboratory for OptoelectronicsAdvanced Biomedical Imaging FacilityHuazhong University of Science and TechnologyWuhan 430074China

Ischemic stroke causes long-term disability and results in motor impairments.Such impairments are associated with structural changes in the neuromuscular junction(NMJ),including detailed morphology and three-dimensional(3D)distribution.However,previous studies only explored morphological changes of individual NMJs after stroke,which limits the understanding of their role in post-stroke motor impairment.Here,we examine 3D distributions and detailed morphology of NMJs in entire mouse muscles after unilateral and bilateral strokes induced by photothrombosis.The results show that 3D distributions and numbers of NMJs do not change after stroke,and severe unilateral stroke causes similar levels of NMJ fragmentation and area enlargement to bilateral stroke.This research provides structural data,deepening the understanding of neuromuscular pathophysiology after stroke.

关键词： tissue optical clearing optical imaging stroke neuromuscular junction

Fluorescence molecular tomography system for in vivo tumor imaging in small animals

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Chinese Optics Letters 2010年第11期8卷 1075-1078页

作者：傅建伟杨孝全全国涛龚辉 Britton Chance Center for Biomedical Photonics Wuhan National Laboratory for OptoelectronicsHuazhong University of Science and Technology

A fluorescence molecular tomography system for in vivo tumor imaging is developed using a 748-nm continuous wave diode laser as an excitation source.A high sensitivity cooled charge-coupled device （CCD） camera with excitation and emission filters is utilized to obtain the excitation and fluorescence images.The laser beam performs fast raster scanning using a dual-axis galvanometric scanner.The accuracy of the laser spot position at the source window is within ±200 μm.Based on the phantom experimental results,the spatial resolution is less than 1.7 mm,and the relative quantitation error is about 10%.In vivo imaging of a tumor-bearing nude mouse tagged with near-infrared dye demonstrates the feasibility of the system.

关键词： Animals Fluorescence Tomography Tumors

Visualization of reticulophagy in living cells using an endoplasmic reticulum-targeted p62 mutant

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Science China(Life Sciences) 2017年第4期60卷 333-344页

作者： Liang Wang Britton Chance Center for Biomedical Photonics Wuhan National Laboratory for Optoelectronics Huazhong University of Science and Technology Wuhan 430074 China MoE Key Laboratory for biomedical photonics Department of Biomedical Engineering Huazhong University of Science and Technology Wuhan 430074 China

Reticulophagy is a type of selective autophagy in which protein aggregate-containing and/or damaged endoplasmic reticulum(ER)fragments are engulfed for lysosomal degradation, which is important for ER homeostasis. Several chemical drugs and mutant proteins that promote protein aggregate formation within the ER lumen can efficiently induce reticulophagy in mammalian cells.However, the exact mechanism and cellular localization of reticulophagy remain unclear. In this report, we took advantage of the self-oligomerization property of p62/SQSTM1, an adaptor for selective autophagy, and developed a novel reticulophagy system based on an ER-targeted p62 mutant to investigate the process of reticulophagy in living cells. LC3 conversion analysis via western blot suggested that p62 mutant aggregate-induced ER stress triggered a cellular autophagic response. Confocal imaging showed that in cells with moderate aggregation conditions, the aggregates of ER-targeted p62 mutants were efficiently sequestered by autophagosomes, which was characterized by colocalization with the autophagosome precursor marker ATG16L1, the omegasome marker DFCP1, and the late autophagosomal marker LC3/GATE-16. Moreover, time-lapse imaging data demonstrated that the LC3-or DFCP1-positive protein aggregates are tightly associated with the reticular structures of the ER, thereby suggesting that reticulophagy occurs at the ER and that omegasomes may be involved in this process.

关键词： reticulophagy 蛋白质总数 endoplasmic 蜂窝胃(嗯) LC3 p62 autophagosomes omegasomes