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      Compact, Fast Blinking Cd-Free Quantum Dots for Super-Resolution Fluorescence Imaging

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          Abstract

          Quantum dots (QDs) can be used as fluorescent probes in single molecule localization microscopy to achieve subdiffraction limit resolution (super-resolution fluorescence imaging). However, the toxicity of Cd in the prototypical CdSe-based QDs can limit their use in biological applications. Furthermore, commercial CdSe QDs are usually modified with relatively thick shells of both inorganic and organic materials to render them in the 10–20 nm size range, which is relatively large for biological labels. In this report, we present compact (4–6 nm) CuInS 2/ZnS (CIS/ZnS) and compare them to commercially sourced CdSe/ZnS QDs for their blinking behavior, localization precision and super-resolution imaging. Although commercial CdSe/ZnS QDs are brighter than the more compact Cd-free CIS/ZnS QD, both give comparable results of 4.5–5.0-fold improvement in imaging resolution over conventional TIRF imaging of actin filaments. This likely results from the fact that CIS/ZnS QDs show very short on-times and long off times which leads to less overlap in the point spread functions of emitting CIS/ZnS QD labels on the actin filaments at the same labeling density. These results demonstrate that CIS/ZnS QDs are an excellent candidate to complement and perhaps even replace the larger and more toxic CdSe-based QDs for robust single- molecule super-resolution imaging.

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          Most cited references53

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          Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).

          We have developed a high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores. In each imaging cycle, only a fraction of the fluorophores were turned on, allowing their positions to be determined with nanometer accuracy. The fluorophore positions obtained from a series of imaging cycles were used to reconstruct the overall image. We demonstrated an imaging resolution of 20 nm. This technique can, in principle, reach molecular-scale resolution.
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            Imaging intracellular fluorescent proteins at nanometer resolution.

            We introduce a method for optically imaging intracellular proteins at nanometer spatial resolution. Numerous sparse subsets of photoactivatable fluorescent protein molecules were activated, localized (to approximately 2 to 25 nanometers), and then bleached. The aggregate position information from all subsets was then assembled into a superresolution image. We used this method--termed photoactivated localization microscopy--to image specific target proteins in thin sections of lysosomes and mitochondria; in fixed whole cells, we imaged vinculin at focal adhesions, actin within a lamellipodium, and the distribution of the retroviral protein Gag at the plasma membrane.
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              Quantum dot bioconjugates for ultrasensitive nonisotopic detection.

              W Chan, S Nie (1998)
              Highly luminescent semiconductor quantum dots (zinc sulfide-capped cadmium selenide) have been covalently coupled to biomolecules for use in ultrasensitive biological detection. In comparison with organic dyes such as rhodamine, this class of luminescent labels is 20 times as bright, 100 times as stable against photobleaching, and one-third as wide in spectral linewidth. These nanometer-sized conjugates are water-soluble and biocompatible. Quantum dots that were labeled with the protein transferrin underwent receptor-mediated endocytosis in cultured HeLa cells, and those dots that were labeled with immunomolecules recognized specific antibodies or antigens.
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                Author and article information

                Journal
                Chem Biomed Eng
                Chem Biomed Eng
                im
                cbihbp
                Chemical & Biomedical Imaging
                Nanjing University and American Chemical Society
                2832-3637
                03 April 2023
                26 June 2023
                : 1
                : 3
                : 251-259
                Affiliations
                []Department of Chemistry and Biochemistry, University of Arkansas , 345 North Campus Drive, Fayetteville, Arkansas 72701, United States
                []Department of Physics, University of Arkansas , 825 West Dickson Street, Fayetteville, Arkansas 72701, United States
                Author notes
                Author information
                https://orcid.org/0000-0002-8601-8302
                https://orcid.org/0000-0003-1124-9947
                Article
                10.1021/cbmi.3c00018
                10302876
                a5b46f5d-336b-4c79-93f7-1b18c9b64820
                © 2023 The Authors. Co-published by Nanjing University and American Chemical Society

                Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works ( https://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 24 January 2023
                : 21 March 2023
                : 20 March 2023
                Funding
                Funded by: National Science Foundation, doi 10.13039/100000001;
                Award ID: CHE-1255440
                Funded by: Arkansas Biosciences Institute, doi 10.13039/100008231;
                Award ID: NA
                Funded by: National Center for Research Resources, doi 10.13039/100000097;
                Award ID: COBRE P30 GM103450
                Categories
                Article
                Custom metadata
                im3c00018
                im3c00018

                copper indium sulfide,zinc sulfide,single molecule,localization microscopy,actin filaments,streptavidin−biotin

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