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Selective labeling of individual neurons in dense cultured networks with nanoparticle-enhanced photoporation

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Abstract
Neurodevelopmental and neurodegenerative disorders are characterized by subtle alterations in synaptic connections and perturbed neuronal network functionality. A hallmark of neuronal connectivity is the presence of dendritic spines, micron-sized protrusions of the dendritic shaft that compartmentalize single synapses to fine-tune synaptic strength. However, accurate quantification of spine density and morphology in mature neuronal networks is hampered by the lack of targeted labeling strategies. To resolve this, we have optimized a method to deliver cell-impermeable compounds into selected cells based on Spatially resolved NAnoparticle-enhanced Photoporation (SNAP). We show that SNAP enables efficient labeling of selected individual neurons and their spines in dense cultured networks without affecting short-term viability. We compare SNAP with widely used spine labeling techniques such as the application of lipophilic dyes and genetically encoded fluorescent markers. Using SNAP, we demonstrate a time-dependent increase in spine density in healthy cultures as well as a reduction in spine density after chemical mimicry of hypoxia. Since the sparse labeling procedure can be automated using an intelligent acquisition scheme, SNAP holds promise for high-content screening campaigns of neuronal connectivity in the context of neurodevelopmental and neurodegenerative disorders.
Keywords
dendritic spine, gold nanoparticle, photoporation, primary neuronal culture, neuron labeling, SNAP, DENDRITIC SPINE DEVELOPMENT, LIVE CELLS, ACTIN, MICROINJECTION, CONNECTIVITY, TRANSMISSION, TRANSFECTION, NANOBUBBLES, PLASTICITY, PHALLOIDIN

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MLA
Xiong, Ranhua, Peter Verstraelen, Jo Demeester, et al. “Selective Labeling of Individual Neurons in Dense Cultured Networks with Nanoparticle-enhanced Photoporation.” FRONTIERS IN CELLULAR NEUROSCIENCE 12 (2018): n. pag. Print.
APA
Xiong, R., Verstraelen, P., Demeester, J., Skirtach, A., Timmermans, J.-P., De Smedt, S., De Vos, W., et al. (2018). Selective labeling of individual neurons in dense cultured networks with nanoparticle-enhanced photoporation. FRONTIERS IN CELLULAR NEUROSCIENCE, 12.
Chicago author-date
Xiong, Ranhua, Peter Verstraelen, Jo Demeester, Andre Skirtach, Jean-Pierre Timmermans, Stefaan De Smedt, Winnok De Vos, and Kevin Braeckmans. 2018. “Selective Labeling of Individual Neurons in Dense Cultured Networks with Nanoparticle-enhanced Photoporation.” Frontiers in Cellular Neuroscience 12.
Chicago author-date (all authors)
Xiong, Ranhua, Peter Verstraelen, Jo Demeester, Andre Skirtach, Jean-Pierre Timmermans, Stefaan De Smedt, Winnok De Vos, and Kevin Braeckmans. 2018. “Selective Labeling of Individual Neurons in Dense Cultured Networks with Nanoparticle-enhanced Photoporation.” Frontiers in Cellular Neuroscience 12.
Vancouver
1.
Xiong R, Verstraelen P, Demeester J, Skirtach A, Timmermans J-P, De Smedt S, et al. Selective labeling of individual neurons in dense cultured networks with nanoparticle-enhanced photoporation. FRONTIERS IN CELLULAR NEUROSCIENCE. 2018;12.
IEEE
[1]
R. Xiong et al., “Selective labeling of individual neurons in dense cultured networks with nanoparticle-enhanced photoporation,” FRONTIERS IN CELLULAR NEUROSCIENCE, vol. 12, 2018.
@article{8572060,
  abstract     = {Neurodevelopmental and neurodegenerative disorders are characterized by subtle alterations in synaptic connections and perturbed neuronal network functionality. A hallmark of neuronal connectivity is the presence of dendritic spines, micron-sized protrusions of the dendritic shaft that compartmentalize single synapses to fine-tune synaptic strength. However, accurate quantification of spine density and morphology in mature neuronal networks is hampered by the lack of targeted labeling strategies. To resolve this, we have optimized a method to deliver cell-impermeable compounds into selected cells based on Spatially resolved NAnoparticle-enhanced Photoporation (SNAP). We show that SNAP enables efficient labeling of selected individual neurons and their spines in dense cultured networks without affecting short-term viability. We compare SNAP with widely used spine labeling techniques such as the application of lipophilic dyes and genetically encoded fluorescent markers. Using SNAP, we demonstrate a time-dependent increase in spine density in healthy cultures as well as a reduction in spine density after chemical mimicry of hypoxia. Since the sparse labeling procedure can be automated using an intelligent acquisition scheme, SNAP holds promise for high-content screening campaigns of neuronal connectivity in the context of neurodevelopmental and neurodegenerative disorders.},
  articleno    = {80},
  author       = {Xiong, Ranhua and Verstraelen, Peter and Demeester, Jo and Skirtach, Andre and Timmermans, Jean-Pierre and De Smedt, Stefaan and De Vos, Winnok and Braeckmans, Kevin},
  issn         = {1662-5102},
  journal      = {FRONTIERS IN CELLULAR NEUROSCIENCE},
  keywords     = {dendritic spine,gold nanoparticle,photoporation,primary neuronal culture,neuron labeling,SNAP,DENDRITIC SPINE DEVELOPMENT,LIVE CELLS,ACTIN,MICROINJECTION,CONNECTIVITY,TRANSMISSION,TRANSFECTION,NANOBUBBLES,PLASTICITY,PHALLOIDIN},
  language     = {eng},
  pages        = {11},
  title        = {Selective labeling of individual neurons in dense cultured networks with nanoparticle-enhanced photoporation},
  url          = {http://dx.doi.org/10.3389/fncel.2018.00080},
  volume       = {12},
  year         = {2018},
}

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