Chemical & Biomedical Imaging最新文献

{"title":"Application of Super-resolution SPEED Microscopy in the Study of Cellular Dynamics","authors":"Wenlan Yu,&nbsp;Coby Rush,&nbsp;Mark Tingey,&nbsp;Samuel Junod and Weidong Yang*,&nbsp;","doi":"10.1021/cbmi.3c00036","DOIUrl":"10.1021/cbmi.3c00036","url":null,"abstract":"<p >Super-resolution imaging techniques have broken the diffraction-limited resolution of light microscopy. However, acquiring three-dimensional (3D) super-resolution information about structures and dynamic processes in live cells at high speed remains challenging. Recently, the development of high-speed single-point edge-excitation subdiffraction (SPEED) microscopy, along with its 2D-to-3D transformation algorithm, provides a practical and effective approach to achieving 3D subdiffraction-limit information in subcellular structures and organelles with rotational symmetry. One of the major benefits of SPEED microscopy is that it does not rely on complex optical components and can be implemented on a standard, inverted epifluorescence microscope, simplifying the process of sample preparation and the expertise requirement. SPEED microscopy is specifically designed to obtain 2D spatial locations of individual immobile or moving fluorescent molecules inside submicrometer biological channels or cavities at high spatiotemporal resolution. The collected data are then subjected to postlocalization 2D-to-3D transformation to obtain 3D super-resolution structural and dynamic information. In recent years, SPEED microscopy has provided significant insights into nucleocytoplasmic transport across the nuclear pore complex (NPC) and cytoplasm-cilium trafficking through the ciliary transition zone. This Review focuses on the applications of SPEED microscopy in studying the structure and function of nuclear pores.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/ce/6f/im3c00036.PMC10369678.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9884011","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Reversible Dual-Channel Near-Infrared Flavonoid Probe for in Vivo Tracking Glutathione Dynamics in Living Mice","authors":"Zijun Zhao,&nbsp;Yutao Zhang*,&nbsp;Menglan Wu,&nbsp;Chenxu Yan and Zhiqian Guo*,&nbsp;","doi":"10.1021/cbmi.3c00051","DOIUrl":"https://doi.org/10.1021/cbmi.3c00051","url":null,"abstract":"<p >Glutathione (GSH) plays a critical role in various biological processes maintaining oxidative homeostasis. However, current reversible probe fluorescence emission is usually in the visible region, making it difficult to monitor glutathione levels in deep tissues and <i>in vivo</i>. Here, we developed a reversible near-infrared fluorescence probe, Flav-N, for real-time tracking of GSH in cells and tissues, which undergoes fast and reversible Michael addition reactions with biothiols. This Flav-N probe showed a rapid and reversible response with GSH at a time of less than 5 s (<i>k</i> = 1286 M^–1S^–1, <i>t</i>_1/2 = 729 ms). Notably, the dynamic changes in the ratio of Flav-N emission intensity at 505 and 728 nm were able to provide real-time feedback on the fluctuation of GSH concentration. We demonstrated that Flav-N enables the performance of fast and reversible imaging of intracellular GSH changes. Importantly, in light of the near-infrared emission and rapid response ability, Flav-N was successfully applied to track GSH dynamics in living mice. This reversible near-infrared NIR probe realizes advances in deep insight into the function of endogenous GSH.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/cbmi.3c00051","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67733089","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fluorescence Lifetime Multiplex Imaging in Expansion Microscopy with Tunable Donor–Acceptor Polymer Dots","authors":"Jie Liu,&nbsp;Zhihe Liu,&nbsp;Feixue Mi,&nbsp;Zihan Yao,&nbsp;Xiaofeng Fang,&nbsp;Yingjie Wang,&nbsp;Zhongying Zhao* and Changfeng Wu*,&nbsp;","doi":"10.1021/cbmi.3c00058","DOIUrl":"https://doi.org/10.1021/cbmi.3c00058","url":null,"abstract":"<p >Fluorescence lifetime imaging microscopy (FLIM) has been widely used in cell biology to detect biomolecules and their interactions. However, breaking the diffraction limit remains a challenge in FLIM due to the typically required photon counting method and the limited photon output of conventional dyes. Here, we introduce semiconducting polymer dots (Pdots) for fluorescence lifetime imaging in expansion microscopy by virtue of their tunable lifetime and huge photon budget. We developed three fluorescent Pdots with average lifetimes ranging from 0.4 to 5 ns by varying the polymer species and compositions. Despite their large spectral overlap, distinctive distributions of the Pdots can be resolved in the lifetime domain. The high fluorescence brightness and large photon output offered by Pdots enable multiplex lifetime imaging in photon-starved expansion microscopy, by which subcellular structures were resolved with a spatial resolution of ∼49 nm. This study reveals the potential of the tunable Pdot probes for lifetime multiplex imaging in expansion microscopy.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/cbmi.3c00058","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49768482","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fluorescent/Phosphorescent Dual-Emissive Probe for Simultaneous and Independent Sensing of Intracellular Hypochlorite and pH via Time-Resolved Luminescence Imaging","authors":"Jun Yang,&nbsp;Meng Wang,&nbsp;Meng Li,&nbsp;Jie Zhou,&nbsp;Kenneth Yin Zhang*,&nbsp;Shujuan Liu and Qiang Zhao*,&nbsp;","doi":"10.1021/cbmi.3c00059","DOIUrl":"10.1021/cbmi.3c00059","url":null,"abstract":"<p >In this work, we present a dual-emissive polymeric probe capable of simultaneous and independent sensing of intracellular hypochlorite and pH. The probe integrates an anthracene–carboxamide-based fluorescent compound with a phosphorescent iridium(III) complex. The fluorescence and phosphorescence spectra are significantly overlapped at 500 to 650 nm, but their lifetimes differ substantially at 14.8 and 1113.6 ns, respectively, thereby allowing for monitoring in different time intervals through time-resolved luminescence analysis. The fluorescence is quenched upon oxidation by hypochlorite, whereas an acidic environment favors protonation of pyridyl groups on the iridium(III) complex, which causes phosphorescence quenching. During hypochlorite sensing, a linear plot of the fluorescence/phosphorescence ratio over hypochlorite concentration was obtained. During pH sensing, the p<i>K</i>_a value was calculated to be about 4.6. The utilization of this probe for imaging cellular endogenous hypochlorite and intracellular pH values was demonstrated. The short-lived and long-lived signals were successfully extracted via time-resolved luminescence imaging microscopy, thereby allowing independent, simultaneous, and quantitative analysis of fluorescence and phosphorescence signals and their response toward hypochlorite and pH values. This study offers potential implications for the development of biosensing and bioimaging applications and paves the way for the simultaneous multiplexed detection of physiological and pathological activities within living cells.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/cbmi.3c00059","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129758579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Circularly Polarized Microscopy of Thin Films of Chiral Organic Dyes","authors":"Andrea Taddeucci,&nbsp;Francesco Zinna,&nbsp;Giuliano Siligardi and Lorenzo Di Bari*,&nbsp;","doi":"10.1021/cbmi.3c00049","DOIUrl":"10.1021/cbmi.3c00049","url":null,"abstract":"<p >We introduce an optical microscopy technique, circularly polarized microscopy or CPM, able to afford spatially resolved electronic circular dichroism (ECD) of thin films of chiral organic semiconductors through a commercial microscope equipped with a camera and inexpensive optics. Provided the dichroic ratio is sufficiently large, the spatial resolution is on the order of the μm and is only limited by the magnification optics integrated in the microscope. We apply CPM to thin films of small chiral π-conjugated molecules, which gave rise to ordered aggregates in the thin layer. Primarily, conventional ECD can reveal and characterize chiral supramolecular structures and possible interferences between anisotropic properties of solid samples; however, it cannot generally account for the spatial distribution of such properties. CPM offers a characterization of supramolecular chirality and of commingling polarization anisotropies of the material, describing their local distribution. To validate CPM, we demonstrated that it can be adopted to quantify the local ECD of samples characterized by intense signals, virtually on any standard optical microscope.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/3b/fc/im3c00049.PMC10467535.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10135171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recognition of Single Fluorescence Events by Temporal Pixel Intensity Fluctuation","authors":"Kai Gu,&nbsp; and ,&nbsp;Chunming Liu*,&nbsp;","doi":"10.1021/cbmi.3c00043","DOIUrl":"https://doi.org/10.1021/cbmi.3c00043","url":null,"abstract":"<p >Single-molecule localization microscopy circumvents the diffraction limit of traditional fluorescence microscopy by detecting the photoemission signals of individual fluorescent molecules. The accurate recognitions of fluorescence molecules/events are critical to single-molecule/super-resolution imaging experiments, which determine the precision of molecular localizations and the quality of the image reconstruction. Herein, we presented a single-molecule detection method which relied on the temporal pixel intensity fluctuation. The method was capable of quickly determining the approximate localizations of fluorescence events with high sensitivity. We evaluated the performance of the method under a series of signal-to-noise ratios (SNR) and discussed the criterion of setting the temporal fluctuation threshold to achieve the optimized spots recognition results.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/cbmi.3c00043","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49768425","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering Isomeric AIEgens Containing Tetraphenylpyrazine for Dual Memory Storage","authors":"Zicheng Liu,&nbsp;Wenhao Wang,&nbsp;Hongfei Liao,&nbsp;Runfeng Lin,&nbsp;Xiang He,&nbsp;Canze Zheng,&nbsp;Changsheng Guo,&nbsp;Hongguang Liu*,&nbsp;Hai-Tao Feng and Ming Chen*,&nbsp;","doi":"10.1021/cbmi.3c00048","DOIUrl":"10.1021/cbmi.3c00048","url":null,"abstract":"<p >Tetraphenylpyrazine (TPP) is a promising heterocycle-based aggregation-induced emission luminogen (AIEgen) which has sparked multiple applications in organic light-emitting diodes, sensors, and biotherapy. However, the utility of it in developing information storage materials is relatively rare. Moreover, TPP is mostly employed as an electronic acceptor in molecular design, while the consideration of it as an electronic donor is attractive in studies which may provide a full understanding of its property to tailor the materials. In this work, we synthesize three TPP-based molecules by decorating it with acrylonitrile and isomeric pyridine units, which show AIE behavior by property inheritance from their parent unit. Interestingly, the effective intramolecular charge transfer takes place from the TPP electronic donor to the acrylonitrile and pyridine electronic acceptor, therefore inducing a remarkable solvatochromic effect as the solvent polarity improves. Moreover, it is revealed that the isomeric effect of the nitrogen atom in the pyridines may pose an influence on the absorption, solvatochromism, and AIE behavior. In addition, the acrylonitrile and pyridine groups are reactive to light and acid–base stimuli with irreversible and reversible responses, respectively. Combined with the high light-harvesting ability of these AIEgens, they show great potential in the stimuli-responsive materials for dual information storage.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/cbmi.3c00048","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127070373","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fluorescence Imaging of Inflammation with Optical Probes","authors":"Cheng Li,&nbsp;Sensen Zhou,&nbsp;Jian Chen and Xiqun Jiang*,&nbsp;","doi":"10.1021/cbmi.3c00039","DOIUrl":"https://doi.org/10.1021/cbmi.3c00039","url":null,"abstract":"<p >Inflammation plays an important role in the occurrence and development of disease; dysregulation of inflammatory progression often leads to disease such as tissue sclerosis, cancers, stroke, etc. Optical imaging technology, due to its higher sensitivity and resolution, can provide finer images for the observation of inflammation. Many optical probes have been developed as contrast agents for optical imaging techniques in different diseases. In this review, we summarize the recent advances of optical probe and imaging methods for imaging inflammation in different organs, such as brain, liver, lung, kidney, intestine, etc. Finally, we discuss the opportunities and challenges of optical probes used in the clinic for inflammation monitoring and prospect their future development in disease detection.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/cbmi.3c00039","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49768689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fluorogenic Reactions in Chemical Biology: Seeing Chemistry in Cells","authors":"Yanyan Chen,&nbsp;Hao Jiang,&nbsp;Tingting Hao,&nbsp;Nan Zhang,&nbsp;Mingyu Li,&nbsp;Xingyun Wang,&nbsp;Xiuxiu Wang*,&nbsp;Wei Wei* and Jing Zhao*,&nbsp;","doi":"10.1021/cbmi.3c00029","DOIUrl":"https://doi.org/10.1021/cbmi.3c00029","url":null,"abstract":"<p >Among the recent development of click chemistry and bioorthogonal chemistry, fluorogenic reactions occupy a unique place in that fluorescence is generated from nonfluorescent reactants, thereby rendering them highly useful and convenient in no-wash live-cell imaging. This topic was extensively reviewed in 2010 by Wang et al. ( <cite><i>Chem. Soc. Rev.</i></cite> <span>2010</span>, <em>39</em>, 1233−1239) and in 2014 by Lin et al. ( <cite><i>Curr. Opin. Chem. Biol.</i></cite> <span>2014</span>, <em>21</em>, 89−95). This review presents a comprehensive and up-to-date overview on the fluorogenic reactions in the past decade. The reactions are classified into four major categories on the basis of the mechanisms of fluorescence generation. Representative examples of each type are discussed briefly in terms of structure, mechanism, and advantages. We describe the latest applications of fluorogenic reactions in chemical biology. In the end, future opportunities and challenges in this field are tentatively proposed.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-05-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/cbmi.3c00029","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"67732810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FRET Imaging of Nonuniformly Distributed DNA SAMs on Gold Reveals the Role Played by the Donor/Acceptor Ratio and the Local Environment in Measuring the Rate of Hybridization","authors":"Adrian Jan Grzędowski,&nbsp;Tianxiao Ma and Dan Bizzotto*,&nbsp;","doi":"10.1021/cbmi.3c00031","DOIUrl":"10.1021/cbmi.3c00031","url":null,"abstract":"<p >Mixed DNA SAMs labeled with a fluorophore (either AlexaFluor488 or AlexaFluor647) were prepared on a single crystal gold bead electrode using potential-assisted thiol exchange and studied using Förster resonance energy transfer (FRET). A measure of the local environment of the DNA SAM (e.g., crowding) was possible using FRET imaging on these surfaces since electrodes prepared this way have a range of surface densities (Γ_DNA). The FRET signal was strongly dependent on Γ_DNA and on the ratio of AlexaFluor488 to AlexaFluor647 used to make the DNA SAM, which were consistent with a model of FRET in 2D systems. FRET was shown to provide a direct measure of the local DNA SAM arrangement on each crystallographic region of interest providing a direct assessment of the probe environment and its influence on the rate of hybridization. The kinetics of duplex formation for these DNA SAMs was also studied using FRET imaging over a range of coverages and DNA SAM compositions. Hybridization of the surface-bound DNA increased the average distance between the fluorophore label and the gold electrode surface and decreased the distance between the donor (D) and acceptor (A), both of which result in an increase in FRET intensity. This increase in FRET was modeled using a second order Langmuir adsorption rate equation, reflecting the fact that both D and A labeled DNA are required to become hybridized to observe a FRET signal. The self-consistent analysis of the hybridization rates on low and high coverage regions on the same electrode showed that the low coverage regions achieved full hybridization 5× faster than the higher coverage regions, approaching rates typically found in solution. The relative increase in FRET intensity from each region of interest was controlled by manipulating the donor to acceptor composition of the DNA SAM without changing the rate of hybridization. The FRET response can be optimized by controlling the coverage and the composition of the DNA SAM sensor surface and could be further improved with the use of a FRET pair with a larger (e.g., &gt; 5 nm) Förster radius.</p>","PeriodicalId":53181,"journal":{"name":"Chemical & Biomedical Imaging","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://ftp.ncbi.nlm.nih.gov/pub/pmc/oa_pdf/1b/a0/im3c00031.PMC10302881.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9741968","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}