Isolating single primary rat hippocampal neurons &amp; astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

Unsworth, Charles P.; Delivopoulos, Evangelos; Gillespie, Trudi; Murray, Alan F.

doi:10.1016/j.biomaterials.2010.12.017

Cited by 27 publications

(22 citation statements)

References 24 publications

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“…It is also highly versatile, as the properties of Parylene-C can be altered via UV irradiation, modifying the permittivity of the substrate to neurons and glia [20]. Moreover, the thickness of Parylene-C can be reduced down to 10 nm rendering this an ideal encapsulant of passive electrodes in capacitive coupling recordings [21]. Finally, our patterning technique can be applied to other cell types, such as the human teratocarcinoma cell line [22], to a single cell resolution [23].…”

Section: Introductionmentioning

confidence: 99%

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

Delivopoulos

Murray²

2011

PLoS ONE

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This paper explores the long term development of networks of glia and neurons on patterns of Parylene-C on a SiO2 substrate. We harvested glia and neurons from the Sprague-Dawley (P1–P7) rat hippocampus and utilized an established cell patterning technique in order to investigate cellular migration, over the course of 3 weeks. This work demonstrates that uncontrolled glial mitosis gradually disrupts cellular patterns that are established early during culture. This effect is not attributed to a loss of protein from the Parylene-C surface, as nitrogen levels on the substrate remain stable over 3 weeks. The inclusion of the anti-mitotic cytarabine (Ara-C) in the culture medium moderates glial division and thus, adequately preserves initial glial and neuronal conformity to underlying patterns. Neuronal apoptosis, often associated with the use of Ara-C, is mitigated by the addition of brain derived neurotrophic factor (BDNF). We believe that with the right combination of glial inhibitors and neuronal promoters, the Parylene-C based cell patterning method can generate structured, active neural networks that can be sustained and investigated over extended periods of time. To our knowledge this is the first report on the concurrent application of Ara-C and BDNF on patterned cell cultures.

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Section: Introductionmentioning

confidence: 99%

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

Delivopoulos

Murray²

2011

PLoS ONE

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“…During their development, astrocytes often migrate very long distances before `settling down' in a specific neural area [34]. To study astrocyte migration, our group has initiated research on how they can be guided to artificial structures [35,36,37]. In this study, we used a phase-contrast live cell imaging system with wide field of view (low magnification) to be able to track human hNT astrocytes moving over relatively long distances (up to several millimeters).…”

Section: Introductionmentioning

confidence: 99%

A Cell Derived Active Contour (CDAC) Method for Robust Tracking in Low Frame Rate, Low Contrast Phase Microscopy - an Example: The Human hNT Astrocyte

2013

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The problem of automated segmenting and tracking of the outlines of cells in microscope images is the subject of active research. While great progress has been made on recognizing cells that are of high contrast and of predictable shape, many situations arise in practice where these properties do not exist and thus many interesting potential studies - such as the migration patterns of astrocytes to scratch wounds - have been relegated to being largely qualitative in nature. Here we analyse a select number of recent developments in this area, and offer an algorithm based on parametric active contours and formulated by taking into account cell movement dynamics. This Cell-Derived Active Contour (CDAC) method is compared with two state-of-the-art segmentation methods for phase-contrast microscopy. Specifically, we tackle a very difficult segmentation problem: human astrocytes that are very large, thin, and irregularly-shaped. We demonstrate quantitatively better results for CDAC as compared to similar segmentation methods, and we also demonstrate the reliable segmentation of qualitatively different data sets that were not possible using existing methods. We believe this new method will enable new and improved automatic cell migration and movement studies to be made.

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“…Cell patterning occurred along the parylene-C strips as expected from [5][6][7][8] due to the surface chemistry between adsorbed fetal bovine serum (FBS) and the parylene-C. However, it should be noted that to avoid a gap between the 50x50μm square ablations the pulse overlap of the laser was set to the smallest distance available of 5μm.…”

Section: B Ir Laser Ablative Patterningmentioning

confidence: 99%

Low cost, patterning of human hNT brain cells on parylene-C with UV & IR laser machining

Raos

Unsworth

Costa

et al. 2013

2013 35th Annual International Conference of the IEEE Engineering in Medicine and Biology Society (EMBC)

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This paper describes the use of 800nm femtosecond infrared (IR) and 248nm nanosecond ultraviolet (UV) laser radiation in performing ablative micromachining of parylene-C on SiO2 substrates for the patterning of human hNT astrocytes. Results are presented that support the validity of using IR laser ablative micromachining for patterning human hNT astrocytes cells while UV laser radiation produces photo-oxidation of the parylene-C and destroys cell patterning. The findings demonstrate how IR laser ablative micromachining of parylene-C on SiO2 substrates can offer a low cost, accessible alternative for rapid prototyping, high yield cell patterning.

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Isolating single primary rat hippocampal neurons & astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

Cited by 27 publications

References 24 publications

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

A Cell Derived Active Contour (CDAC) Method for Robust Tracking in Low Frame Rate, Low Contrast Phase Microscopy - an Example: The Human hNT Astrocyte

Low cost, patterning of human hNT brain cells on parylene-C with UV & IR laser machining

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Isolating single primary rat hippocampal neurons & astrocytes on ultra-thin patterned parylene-C/silicon dioxide substrates

Cited by 27 publications

References 24 publications

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

Controlled Adhesion and Growth of Long Term Glial and Neuronal Cultures on Parylene-C

A Cell Derived Active Contour (CDAC) Method for Robust Tracking in Low Frame Rate, Low Contrast Phase Microscopy - an Example: The Human hNT Astrocyte

Low cost, patterning of human hNT brain cells on parylene-C with UV &amp; IR laser machining

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Product

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Low cost, patterning of human hNT brain cells on parylene-C with UV & IR laser machining