Dataset Information

Dual-Color-Emitting Carbon Nanodots for Multicolor Bioimaging and Optogenetic Control of Ion Channels.

ABSTRACT: The development of intrinsically multicolor-emitting carbon nanodots (CNDs) has been one of the great challenges for their various fields of applications. Here, the controlled electronic structure engineering of CNDs is performed to emit two distinct colors via the facile surface modification with 4-octyloxyaniline. The so-called dual-color-emitting CNDs (DC-CNDs) can be stably encapsulated within poly(styrene-co-maleic anhydride) (PSMA). The prepared water-soluble DC-CNDs@PSMA can be successfully applied to in vitro and in vivo dual-color bioimaging and optogenetics. In vivo optical imaging can visualize the biodistribution of intravenously injected DC-CNDs@PSMA. In addition, the light-triggered activation of ion channel, channelrhodopsin-2, for optogenetic applications is demonstrated. As a new type of fluorophore, DC-CNDs offer a big insight into the design of charge-transfer complexes for various optical and biomedical applications.

SUBMITTER: Kim H

PROVIDER: S-EPMC5700631 | biostudies-other | 2017 Nov

REPOSITORIES: biostudies-other

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Dual-Color-Emitting Carbon Nanodots for Multicolor Bioimaging and Optogenetic Control of Ion Channels.

Kim Hyemin H Park Yoonsang Y Beack Songeun S Han Seulgi S Jung Dooyup D Cha Hyung Joon HJ Kwon Woosung W Hahn Sei Kwang SK

Advanced science (Weinheim, Baden-Wurttemberg, Germany) 20171003 11

The development of intrinsically multicolor-emitting carbon nanodots (CNDs) has been one of the great challenges for their various fields of applications. Here, the controlled electronic structure engineering of CNDs is performed to emit two distinct colors via the facile surface modification with 4-octyloxyaniline. The so-called dual-color-emitting CNDs (DC-CNDs) can be stably encapsulated within poly(styrene-<i>co</i>-maleic anhydride) (PSMA). The prepared water-soluble DC-CNDs@PSMA can be suc ...[more]

PMID: 29201627

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Project description:Optogenetic assays provide a flexible, scalable, and information rich approach to probe compound effects for ion channel drug targets in both heterologous expression systems and associated disease relevant cell types. Despite the potential utility and growing adoption of optogenetics, there remains a critical need for compatible platform technologies with the speed, sensitivity, and throughput to enable their application to broader drug screening applications. To address this challenge, we developed the SwarmTM, a custom designed optical instrument for highly parallelized, multicolor measurements in excitable cells, simultaneously recording changes in voltage and calcium activities at high temporal resolution under optical stimulation. The compact design featuring high power LEDs, large numerical aperture optics, and fast photodiode detection enables all-optical individual well readout of 24-wells simultaneously from multi-well plates while maintaining sufficient temporal resolution to probe millisecond response dynamics. The Swarm delivers variable intensity blue-light optogenetic stimulation to enable membrane depolarization and red or lime-light excitation to enable fluorescence detection of the resulting changes in membrane potential or calcium levels, respectively. The Swarm can screen ~10,000 wells/day in 384-well format, probing complex pharmacological interactions via a wide array of stimulation protocols. To evaluate the Swarm screening system, we optimized a series of heterologous optogenetic spiking HEK293 cell assays for several voltage-gated sodium channel subtypes including Nav1.2, Nav1.5, and Nav1.7. The Swarm was able to record pseudo-action potentials stably across all 24 objectives and provided pharmacological characterization of diverse sodium channel blockers. We performed a Nav1.7 screen of 200,000 small molecules in a 384-well plate format with all 560 plates reaching a Z' > 0.5. As a demonstration of the versatility of the Swarm, we also developed an assay measuring cardiac action potential and calcium waveform properties simultaneously under paced conditions using human induced pluripotent stem (iPS) cell-derived cardiomyocytes as an additional counter screen for cardiac toxicity. In summary, the Swarm is a novel high-throughput all-optical system capable of collecting information-dense data from optogenetic assays in both heterologous and iPS cell-derived models, which can be leveraged to drive diverse therapeutic discovery programs for nervous system disorders and other disease areas involving excitable cells.

Dataset Information

Dual-Color-Emitting Carbon Nanodots for Multicolor Bioimaging and Optogenetic Control of Ion Channels.

Publications

Dual-Color-Emitting Carbon Nanodots for Multicolor Bioimaging and Optogenetic Control of Ion Channels.

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