Three-dimensional imaging cannot be achieved easily using previously developed localization super-resolution techniques. Here, we present a three-dimensional multimodal sub-diffraction imaging technique with spinning-...Three-dimensional imaging cannot be achieved easily using previously developed localization super-resolution techniques. Here, we present a three-dimensional multimodal sub-diffraction imaging technique with spinning-disk （SD） confocal microscopy called 3D-MUSIC, which not only has all the advantages of SD confocal microscopy such as fast imaging speed, high signal-to-noise ratio, and optical-sectioning capability, but also extends its spatial resolution limit along all three dimensions. Both axial and lateral resolution can be improved simul- taneously by virtue of the blinking/fluctuating nature of modified fluorescent probes, exemplified with the quantum dots. Further, super-resolution images with dual modality can be obtained through super-resolution optical fluctuation imaging （SOFI） and bleaching/blinking-assisted localization microscopy （BALM）. Therefore, fast super-resolution imaging can be achieved with SD-SOFI by capturing only 100 frames while SD-BaLM yields high-resolution imaging.展开更多
Carbon dots(CDs)have emerged as promising nanomaterials for bioimaging and stress monitoring due to their unique optical and functional properties.CDs were synthesized using citric acid and o-phenylenediamine via micr...Carbon dots(CDs)have emerged as promising nanomaterials for bioimaging and stress monitoring due to their unique optical and functional properties.CDs were synthesized using citric acid and o-phenylenediamine via microwave-assisted heating,named as CP-CDs.High-resolution transmission electron microscopy observed an average particle size of 3.65±0.40 nm with graphitic cores.Raman spectroscopy and Fourier transform infrared spectroscopy confirmed diverse functional groups.The CDs exhibited excitation-dependent fluorescence with a peak emission at 432 nm,a high quantum yield of 54.91%,and a fluorescence lifetime of 9.50±0.15 ns,making them highly suitable for bioimaging.Confocal microscopy demonstrated tissue-specific localization in lettuce plant cells.In stem cells,CP-CDs predominantly targeted mitochondria,confirmed by a colocalization with Mito-Tracker Red.In contrast,leaf cells showed selective accumulation at the stomatal openings.Under salt-and heat-induced stress,stem cells exhibited an increase in mitochondrial fluorescence,indicating stress-responsive interactions,whereas leaf cells maintained consistent stomatal localization.Further,enhanced fluorescence from chloroplasts under stress conditions suggested synergistic effects with chlorophyll.Also,stress conditions caused CP-CDs to accumulate at the cell boundaries in stem cells,highlighting their sensitivity to stress-induced changes.These findings demonstrate the optical properties,tissue-specific uptake,and organelle-level localization of CP-CDs,underlining their potential for bioimaging,stress detection,and targeted delivery systems in plants.展开更多
文摘Three-dimensional imaging cannot be achieved easily using previously developed localization super-resolution techniques. Here, we present a three-dimensional multimodal sub-diffraction imaging technique with spinning-disk （SD） confocal microscopy called 3D-MUSIC, which not only has all the advantages of SD confocal microscopy such as fast imaging speed, high signal-to-noise ratio, and optical-sectioning capability, but also extends its spatial resolution limit along all three dimensions. Both axial and lateral resolution can be improved simul- taneously by virtue of the blinking/fluctuating nature of modified fluorescent probes, exemplified with the quantum dots. Further, super-resolution images with dual modality can be obtained through super-resolution optical fluctuation imaging （SOFI） and bleaching/blinking-assisted localization microscopy （BALM）. Therefore, fast super-resolution imaging can be achieved with SD-SOFI by capturing only 100 frames while SD-BaLM yields high-resolution imaging.
基金supported by the National Natural Science Foundation of China(Grant 52172033)the support of the Key Laboratory of Structure and Functional Regulation of Hybrid Materials of the Ministry of Education,Anhui University,China.
文摘Carbon dots(CDs)have emerged as promising nanomaterials for bioimaging and stress monitoring due to their unique optical and functional properties.CDs were synthesized using citric acid and o-phenylenediamine via microwave-assisted heating,named as CP-CDs.High-resolution transmission electron microscopy observed an average particle size of 3.65±0.40 nm with graphitic cores.Raman spectroscopy and Fourier transform infrared spectroscopy confirmed diverse functional groups.The CDs exhibited excitation-dependent fluorescence with a peak emission at 432 nm,a high quantum yield of 54.91%,and a fluorescence lifetime of 9.50±0.15 ns,making them highly suitable for bioimaging.Confocal microscopy demonstrated tissue-specific localization in lettuce plant cells.In stem cells,CP-CDs predominantly targeted mitochondria,confirmed by a colocalization with Mito-Tracker Red.In contrast,leaf cells showed selective accumulation at the stomatal openings.Under salt-and heat-induced stress,stem cells exhibited an increase in mitochondrial fluorescence,indicating stress-responsive interactions,whereas leaf cells maintained consistent stomatal localization.Further,enhanced fluorescence from chloroplasts under stress conditions suggested synergistic effects with chlorophyll.Also,stress conditions caused CP-CDs to accumulate at the cell boundaries in stem cells,highlighting their sensitivity to stress-induced changes.These findings demonstrate the optical properties,tissue-specific uptake,and organelle-level localization of CP-CDs,underlining their potential for bioimaging,stress detection,and targeted delivery systems in plants.
