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      Current and future advances in fluorescence-based visualization of plant cell wall components and cell wall biosynthetic machineries

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          Abstract

          Plant cell wall-derived biomass serves as a renewable source of energy and materials with increasing importance. The cell walls are biomacromolecular assemblies defined by a fine arrangement of different classes of polysaccharides, proteoglycans, and aromatic polymers and are one of the most complex structures in Nature. One of the most challenging tasks of cell biology and biomass biotechnology research is to image the structure and organization of this complex matrix, as well as to visualize the compartmentalized, multiplayer biosynthetic machineries that build the elaborate cell wall architecture. Better knowledge of the plant cells, cell walls, and whole tissue is essential for bioengineering efforts and for designing efficient strategies of industrial deconstruction of the cell wall-derived biomass and its saccharification. Cell wall-directed molecular probes and analysis by light microscopy, which is capable of imaging with a high level of specificity, little sample processing, and often in real time, are important tools to understand cell wall assemblies. This review provides a comprehensive overview about the possibilities for fluorescence label-based imaging techniques and a variety of probing methods, discussing both well-established and emerging tools. Examples of applications of these tools are provided. We also list and discuss the advantages and limitations of the methods. Specifically, we elaborate on what are the most important considerations when applying a particular technique for plants, the potential for future development, and how the plant cell wall field might be inspired by advances in the biomedical and general cell biology fields.

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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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              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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                Author and article information

                Contributors
                btdevree@plen.ku.dk
                lms@plen.ku.dk
                glaz@plen.ku.dk
                fr@plen.ku.dk
                klaus.herburger@plen.ku.dk
                staffan.persson@plen.ku.dk
                mravec@plen.ku.dk
                Journal
                Biotechnol Biofuels
                Biotechnol Biofuels
                Biotechnology for Biofuels
                BioMed Central (London )
                1754-6834
                29 March 2021
                29 March 2021
                2021
                : 14
                : 78
                Affiliations
                [1 ]GRID grid.5254.6, ISNI 0000 0001 0674 042X, Department of Plant and Environmental Sciences, , University of Copenhagen, ; Thorvaldsensvej 40, 1871 Frederiksberg, Denmark
                [2 ]GRID grid.16821.3c, ISNI 0000 0004 0368 8293, Joint International Research Laboratory of Metabolic and Developmental Sciences, State Key Laboratory of Hybrid Rice, School of Life Sciences and Biotechnology, , Shanghai Jiao Tong University, ; Shanghai, China
                Author information
                http://orcid.org/0000-0003-0582-8125
                http://orcid.org/0000-0002-6868-0423
                http://orcid.org/0000-0002-8257-4432
                http://orcid.org/0000-0002-0856-5511
                http://orcid.org/0000-0001-8014-0844
                http://orcid.org/0000-0002-6377-5132
                http://orcid.org/0000-0003-0331-3304
                Article
                1922
                10.1186/s13068-021-01922-0
                8008654
                33781321
                8647d491-03c4-4529-8773-2925275827ce
                © The Author(s) 2021

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

                History
                : 17 November 2020
                : 5 March 2021
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100004836, Det Frie Forskningsråd;
                Award ID: 272-07-0152
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100008398, Villum Fonden;
                Award ID: 17489
                Award ID: 00023089
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/100008398, Villum Fonden;
                Award ID: 25905
                Award Recipient :
                Funded by: Australian Research Council
                Award ID: DP190101941
                Award ID: FT160100218
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100009708, Novo Nordisk Fonden;
                Award ID: NNF19OC0056076
                Award Recipient :
                Categories
                Review
                Custom metadata
                © The Author(s) 2021

                Biotechnology
                plant cell wall architecture,fluorescence microscopy,superresolution microscopy,live cell imaging,cell wall probes,nanobodies,lambodies,click chemistry,genetic probes,aptamers,carbohydrate-binding modules,anti-glycan antibodies

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