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| Home > Documents in Process > Fluorogenic Chemical Probes for Wash-free Imaging of Cell Membrane Damage in Ferroptosis, Necrosis, and Axon Injury. > print |
| Home > Documents in Process > Fluorogenic Chemical Probes for Wash-free Imaging of Cell Membrane Damage in Ferroptosis, Necrosis, and Axon Injury. > print |
| 001 | 282305 | ||
| 005 | 20251117145937.0 | ||
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| 100 | 1 | _ | |a Mauker, Philipp |b 0 |
| 245 | _ | _ | |a Fluorogenic Chemical Probes for Wash-free Imaging of Cell Membrane Damage in Ferroptosis, Necrosis, and Axon Injury. |
| 260 | _ | _ | |a Washington, DC |c 2024 |b ACS Publications |
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| 520 | _ | _ | |a Selectively labeling cells with damaged membranes is needed not only for identifying dead cells in culture, but also for imaging membrane barrier dysfunction in pathologies in vivo. Most membrane permeability stains are permanently colored or fluorescent dyes that need washing to remove their non-uptaken extracellular background and reach good image contrast. Others are DNA-binding environment-dependent fluorophores, which lack design modularity, have potential toxicity, and can only detect permeabilization of cell volumes containing a nucleus (i.e., cannot delineate damaged volumes in vivo nor image non-nucleated cell types or compartments). Here, we develop modular fluorogenic probes that reveal the whole cytosolic volume of damaged cells, with near-zero background fluorescence so that no washing is needed. We identify a specific disulfonated fluorogenic probe type that only enters cells with damaged membranes, then is enzymatically activated and marks them. The esterase probe MDG1 is a reliable tool to reveal live cells that have been permeabilized by biological, biochemical, or physical membrane damage, and it can be used in multicolor microscopy. We confirm the modularity of this approach by also adapting it for improved hydrolytic stability, as the redox probe MDG2. We conclude by showing the unique performance of MDG probes in revealing axonal membrane damage (which DNA fluorogens cannot achieve) and in discriminating damage on a cell-by-cell basis in embryos in vivo. The MDG design thus provides powerful modular tools for wash-free in vivo imaging of membrane damage, and indicates how designs may be adapted for selective delivery of drug cargoes to these damaged cells: offering an outlook from selective diagnosis toward therapy of membrane-compromised cells in disease. |
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| 700 | 1 | _ | |a Beckmann, Daniela |b 1 |
| 700 | 1 | _ | |a Kitowski, Annabel |b 2 |
| 700 | 1 | _ | |a Heise, Constanze |b 3 |
| 700 | 1 | _ | |a Wientjens, Chantal |b 4 |
| 700 | 1 | _ | |a Davidson, Andrew J |b 5 |
| 700 | 1 | _ | |a Wanderoy, Simone |b 6 |
| 700 | 1 | _ | |a Fabre, Gabin |b 7 |
| 700 | 1 | _ | |a Harbauer, Angelika B |b 8 |
| 700 | 1 | _ | |a Wood, Will |b 9 |
| 700 | 1 | _ | |a Wilhelm, Christoph |b 10 |
| 700 | 1 | _ | |a Thorn-Seshold, Julia |0 0000-0002-4879-4159 |b 11 |
| 700 | 1 | _ | |a Misgeld, Thomas |0 P:(DE-2719)2810727 |b 12 |u dzne |
| 700 | 1 | _ | |a Kerschensteiner, Martin |b 13 |
| 700 | 1 | _ | |a Thorn-Seshold, Oliver |0 0000-0003-3981-651X |b 14 |
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