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  1. Article ; Online: Directionality quantification of in vitro grown dorsal root ganglion neurites using Fast Fourier Transform.

    Emilie, Lerognon / Tihana, Marciuš / Antonio, Šarolić / Damir, Kovačić / Dries, Braeken / Damir, Sapunar

    Journal of neuroscience methods

    2023  Volume 386, Page(s) 109796

    Abstract: Background: The directionality analysis of the neurite outgrowths is an important methodology in neuroscience, especially in determining the behavior of neurons grown on silicon substrates.: New method: Here we aimed to describe the methodology for ... ...

    Abstract Background: The directionality analysis of the neurite outgrowths is an important methodology in neuroscience, especially in determining the behavior of neurons grown on silicon substrates.
    New method: Here we aimed to describe the methodology for quantification of the directionality of neurites based on the Fast Fourier Transform (FFT). We performed an image analysis case study that incorporates several software solutions and provides a rapid and precise technique to determine the directionality of neurites. In order to elicit aligned or unaligned neurite growth patterns, we used adult and newborn dorsal root ganglion (DRG) neurons grown on silicon micro-pillar substrates (MPS) with different pillar widths and spacing.
    Results: Compared to the control glass surfaces the neonatal and adult N52 and IB4 DRG neurites exhibited regular growth patterns more pronounced in the MPS regions with s narrow pillar spacing range. The neurites were preferentially oriented along three directional axes at 30°, 90°, and 150°.
    Conclusion: The proposed methodology showed that FFT analysis is a reliable and easily reproducible method that can be successfully used to test growth patterns of DRG neurites grown on different substrates by considering the direction and angle of the neurites as well as the size of the soma.
    MeSH term(s) Infant, Newborn ; Humans ; Neurites/physiology ; Ganglia, Spinal ; Fourier Analysis ; Silicon ; Cells, Cultured
    Chemical Substances Silicon (Z4152N8IUI)
    Language English
    Publishing date 2023-01-15
    Publishing country Netherlands
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 282721-9
    ISSN 1872-678X ; 0165-0270
    ISSN (online) 1872-678X
    ISSN 0165-0270
    DOI 10.1016/j.jneumeth.2023.109796
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Advantageous environment of micro-patterned, high-density complementary metal–oxide–semiconductor electrode array for spiral ganglion neurons cultured in vitro

    Viktorija Radotić / Dries Braeken / Petar Drviš / Marta Mattotti / Damir Kovačić

    Scientific Reports, Vol 8, Iss 1, Pp 1-

    2018  Volume 19

    Abstract: Abstract This study investigated micro-patterned, high-density complementary metal–oxide–semiconductor (CMOS) electrode array to be used as biologically permissive environment for organization, guidance and electrical stimulation of spiral ganglion ... ...

    Abstract Abstract This study investigated micro-patterned, high-density complementary metal–oxide–semiconductor (CMOS) electrode array to be used as biologically permissive environment for organization, guidance and electrical stimulation of spiral ganglion neurons (SGN). SGNs extracted and isolated from cochleae of P5-P7 rat pups and adult guinea pigs were cultured 1, 4 and 7 days in vitro on glass coverslips (control) and CMOS electrode array. The cultures were analyzed visually and immunohistochemically for SGN presence, outgrowth, neurite alignment, neurite length, neurite asymmetry as well as the contact of a neuronal soma and neurites with the micro-electrodes. Our findings indicate that topographical environment of CMOS chip with micro-patterned pillars enhanced growth, survival, morphology, neural orientation and alignment of SGNs in vitro compared to control. Smaller spacing (0.8–1.6 µm) between protruding pillars on CMOS led SGNs to develop structured and guided neurites oriented along three topographical axes separated by 60°. We found morphological basis for positioning of the micro-electrodes on the chip that was appropriate for direct contact of SGNs with them. This configuration allowed CMOS electrode array to electrically stimulate the SGN whose responses were observed with live Fluo 4 calcium imaging.
    Keywords Medicine ; R ; Science ; Q
    Subject code 620
    Language English
    Publishing date 2018-05-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Substrate topography determines neuronal polarization and growth in vitro.

    Liesbeth Micholt / Annette Gärtner / Dimiter Prodanov / Dries Braeken / Carlos G Dotti / Carmen Bartic

    PLoS ONE, Vol 8, Iss 6, p e

    2013  Volume 66170

    Abstract: The establishment of neuronal connectivity depends on the correct initial polarization of the young neurons. In vivo, developing neurons sense a multitude of inputs and a great number of molecules are described that affect their outgrowth. In vitro, many ...

    Abstract The establishment of neuronal connectivity depends on the correct initial polarization of the young neurons. In vivo, developing neurons sense a multitude of inputs and a great number of molecules are described that affect their outgrowth. In vitro, many studies have shown the possibility to influence neuronal morphology and growth by biophysical, i.e. topographic, signaling. In this work we have taken this approach one step further and investigated the impact of substrate topography in the very early differentiation stages of developing neurons, i.e. when the cell is still at the round stage and when the first neurite is forming. For this purpose we fabricated micron sized pillar structures with highly reproducible feature sizes, and analyzed neurons on the interface of flat and topographic surfaces. We found that topographic signaling was able to attract the polarization markers of mouse embryonic neurons -N-cadherin, Golgi-centrosome complex and the first bud were oriented towards topographic stimuli. Consecutively, the axon was also preferentially extending along the pillars. These events seemed to occur regardless of pillar dimensions in the range we examined. However, we found differences in neurite length that depended on pillar dimensions. This study is one of the first to describe in detail the very early response of hippocampal neurons to topographic stimuli.
    Keywords Medicine ; R ; Science ; Q
    Subject code 590 ; 571
    Language English
    Publishing date 2013-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article: In vitro recording of neural activity using carbon nanosheet microelectrodes

    Collaert, Nadine / Carolina Mora Lopez / Daire J. Cott / Dries Braeken / Jordi Cools / Michael De Volder

    Carbon. 2014 Feb., v. 67

    2014  

    Abstract: The advent of nanotechnology has revolutionised our ability to engineer electrode interfaces. These are particularly attractive to measure biopotentials, and to study the nervous system. In this work, we demonstrate enhanced in vitro recording of ... ...

    Abstract The advent of nanotechnology has revolutionised our ability to engineer electrode interfaces. These are particularly attractive to measure biopotentials, and to study the nervous system. In this work, we demonstrate enhanced in vitro recording of neuronal activity using electrodes decorated with carbon nanosheets (CNSs). This material comprises of vertically aligned, free standing conductive sheets of only a few graphene layers with a high surface-area-to-volume ratio, which makes them an interesting material for biomedical electrodes. Further, compared to carbon nanotubes, CNSs can be synthesised without the need for metallic catalysts like Ni, Co or Fe, thereby reducing potential cytotoxicity risks. Electrochemical measurements show a five times higher charge storage capacity, and an almost ten times higher double layer capacitance as compared to TiN. In vitro experiments were performed by culturing primary hippocampal neurons from mice on micropatterned electrodes. Neurophysiological recordings exhibited high signal-to-noise ratios of 6.4, which is a twofold improvement over standard TiN electrodes under the same conditions.
    Keywords capacitance ; carbon nanotubes ; catalysts ; cobalt ; cytotoxicity ; electrochemistry ; graphene ; in vitro studies ; iron ; mice ; microelectrodes ; nanosheets ; neurons ; neurophysiology ; nickel ; risk
    Language English
    Dates of publication 2014-02
    Size p. 178-184.
    Publishing place Elsevier Ltd
    Document type Article
    ISSN 0008-6223
    DOI 10.1016/j.carbon.2013.09.079
    Database NAL-Catalogue (AGRICOLA)

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