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      Transforming the development and dissemination of cutting-edge microscopy and computation

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          Abstract

          We propose a network of National Imaging Centers that provide collaborative, interdisciplinary spaces needed for developing, applying and teaching advanced biological imaging techniques. Our proposal is based on recommendations from an NSF sponsored workshop on realizing the promise of innovations in imaging and computation for biological discovery.

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          Most cited references18

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          Content-aware image restoration: pushing the limits of fluorescence microscopy

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            Dual-color superresolution imaging of genetically expressed probes within individual adhesion complexes.

            Accurate determination of the relative positions of proteins within localized regions of the cell is essential for understanding their biological function. Although fluorescent fusion proteins are targeted with molecular precision, the position of these genetically expressed reporters is usually known only to the resolution of conventional optics ( approximately 200 nm). Here, we report the use of two-color photoactivated localization microscopy (PALM) to determine the ultrastructural relationship between different proteins fused to spectrally distinct photoactivatable fluorescent proteins (PA-FPs). The nonperturbative incorporation of these endogenous tags facilitates an imaging resolution in whole, fixed cells of approximately 20-30 nm at acquisition times of 5-30 min. We apply the technique to image different pairs of proteins assembled in adhesion complexes, the central attachment points between the cytoskeleton and the substrate in migrating cells. For several pairs, we find that proteins that seem colocalized when viewed by conventional optics are resolved as distinct interlocking nano-aggregates when imaged via PALM. The simplicity, minimal invasiveness, resolution, and speed of the technique all suggest its potential to directly visualize molecular interactions within cellular structures at the nanometer scale.
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              Teaching Old Dyes New Tricks: Biological Probes Built from Fluoresceins and Rhodamines.

              Small-molecule fluorophores, such as fluorescein and rhodamine derivatives, are critical tools in modern biochemical and biological research. The field of chemical dyes is old; colored molecules were first discovered in the 1800s, and the fluorescein and rhodamine scaffolds have been known for over a century. Nevertheless, there has been a renaissance in using these dyes to create tools for biochemistry and biology. The application of modern chemistry, biochemistry, molecular genetics, and optical physics to these old structures enables and drives the development of novel, sophisticated fluorescent dyes. This critical review focuses on an important example of chemical biology-the melding of old and new chemical knowledge-leading to useful molecules for advanced biochemical and biological experiments.
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                Author and article information

                Journal
                Nature Methods
                Nat Methods
                Springer Science and Business Media LLC
                1548-7091
                1548-7105
                August 2019
                July 30 2019
                August 2019
                : 16
                : 8
                : 667-669
                Article
                10.1038/s41592-019-0475-y
                7643542
                31363203
                e6215836-ad14-471c-843f-0ce4665e951e
                © 2019

                http://www.springer.com/tdm

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