Infra-red laser ablative micromachining of parylene-C on SiO2substrates for rapid prototyping, high yield, human neuronal cell patterning

Cell patterning commonly employs photolithographic methods for the micro fabrication of structures on silicon chips. These require expensive photo-mask development and complex photolithographic processing. Laser based patterning of cells has been studied in vitro and laser ablation of polymers is an active area of research promising high aspect ratios. This paper disseminates how 800 nm femtosecond infrared (IR) laser radiation can be successfully used to perform laser ablative micromachining of parylene-C on SiO2 substrates for the patterning of human hNT astrocytes (derived from the human teratocarcinoma cell line (hNT)) whilst 248 nm nanosecond ultra-violet laser radiation produces photo-oxidization of the parylene-C and destroys cell patterning. In this work, we report the laser ablation methods used and the ablation characteristics of parylene-C for IR pulse fluences. Results follow that support the validity of using IR laser ablative micromachining for patterning human hNT astrocytes cells. We disseminate the variation in yield of patterned hNT astrocytes on parylene-C with laser pulse spacing, pulse number, pulse fluence and parylene-C strip width. The findings demonstrate how laser ablative micromachining of parylene-C on SiO2 substrates can offer an accessible alternative for rapid prototyping, high yield cell patterning with broad application to multi-electrode arrays, cellular micro-arrays and microfluidics.

Patterning and detailed study of human hNT astrocytes on parylene-C/silicon dioxide substrates to the single cell level

It is estimated that the adult human brain contains 100 billion neurons with 5–10 times as many astrocytes. Although it has been generally considered that the astrocyte is a simple supportive cell to the neuron, recent research has revealed new functionality of the astrocyte in the form of information transfer to neurons of the brain. In our previous work we developed a protocol to pattern the hNT neuron (derived from the human teratocarcinoma cell line (hNT)) on parylene-C/SiO2 substrates. In this work, we report how we have managed to pattern hNT astrocytes, on parylene-C/SiO2 substrates to single cell resolution. This article disseminates the nanofabrication and cell culturing steps necessary for the patterning of such cells. In addition, it reports the necessary strip lengths and strip width dimensions of parylene-C that encourage high degrees of cellular coverage and single cell isolation for this cell type. The significance in patterning the hNT astrocyte on silicon chip is that it will help enable single cell and network studies into the undiscovered functionality of this interesting cell, thus, contributing to closer pathological studies of the human brain.

Effects of parylene-C photooxidation on serum-assisted glial and neuronal patterning

The increasing use of patterned neural networks in multielectrode arrays and similar devices drives the constant development and evaluation of new biomaterials. Recently, we presented a promising technique to guide neurons and glia reliably and effectively. Parylene-C, a common hydrophobic polymer, was photolithographically patterned on silicon oxide (SiO2) and subsequently activated via immersion in serum. In this article, we explore the effects of ultraviolet (UV)-induced oxidation on parylene's ability to pattern neurons and glia. We exposed parylene-C stripe patterns to increasing levels of UV radiation and found a dose-dependent reduction in the total mass of patterned cells, as well as a gradual loss of glial and neuronal conformity to the patterns. In contrast, nonirradiated patterns had superior patterning results and increased presence of cells. The reduced cell adhesion and patterning after the formation of aldehyde and carboxyl groups on UV-radiated parylene-C supports our hypothesis that cell adhesion and growth on parylene is facilitated by hydrophobic adsorption of serum proteins. We conclude that unlike other cell patterning schemes, our technique does not rely on photooxidation of the polymer. Nonetheless, the precise control of oxygenated groups on parylene could pave the way for the differential binding of proteins and other molecules on the surface, aiding in the adhesion of alternative cell types. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010

Seal Formation in Silicon Planar Patch-Clamp Microstructures

This paper presents a microfabricated planar patch-clamp electrode design and looks at the impact of several physical characteristics on seal formation. The device consists of a patch aperture, 1.5-2.5 mum in diameter and 7-12 mum in depth, with a reverse-side deep-etched 80-mum well. The patch aperture was coated with either thermal oxide or plasma-enhanced chemical vapor deposited (PECVD) SiO2. Some of the thermal oxide devices were converted into protruding nozzle structures, and some were boron-doped. Seal formation was tested with cultured N2a neuroblastoma cells. The PECVD oxide devices produced an average seal resistance of 34 MOmega(n = 24), and the thermal oxide devices produced an average seal resistance of 96 MOmega(n = 59). Seal resistance was found to positively correlate with patch aperture depth. Whole-cell recordings were obtained from 14% of cells tested with the thermal oxide devices, including a single recording where a gigaohm seal was obtained.

Controlled adhesion and growth of long term glial and neuronal cultures on Parylene-C

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.

First human hNT astrocytes patterned to single cell resolution on parylene-C/Silicon dioxide substrates

In our previous work we developed a successful protocol to pattern the human hNT neuron (derived from the human teratocarcinoma cell line (hNT)) on parylene-C/SiO2 substrates. This communication, reports how we have successfully managed to pattern the supportive cell to the neuron, the hNT astrocyte, on such substrates. Here we disseminate the nanofabrication, cell differentiation and cell culturing protocols necessary to successfully pattern the first human hNT astrocytes to single cell resolution on parylene-C/SiO2 substrates. This is performed for varying parylene strip widths providing excellent contrast to the SiO2 substrate and elegant single cell isolation at 10μm strip widths. The breakthrough in patterning human cells on a silicon chip has widespread implications and is valuable as a platform technology as it enables a detailed study of the human brain at the cellular and network level.

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

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.

All-optical switching device for infrared based on PbTe quantum dots

Multilayers of PbTe quantum dots embedded in SiO2 were fabricated by alternate use of Pulsed Laser Deposition (PLD) and Plasma Enhanced Chemical Vapor Deposition (PECVD) techniques. The morphological properties of the nanostructured material were studied by means of High Resolution Transmission Electron Microscopy (HRTEM), Grazing-Incidence Small-Angle X-ray scattering (GISAXS) and X-ray Reflectometry (XRR) techniques. A preliminary analysis of the GISAXS spectra provided information about the multilayer periodicity and its relationship to the size of the deposited PbTe nanoparticles. Finally multilayers were fabricated inside a Fabry-Perot cavity. The device was characterized by means of Scanning Electron Microscopy (SEM). Transmittance measurements show the device functionality in the infrared region. (C) 2007 Elsevier Ltd. All rights reserved.

TL dosimetry of natural quartz sensitized by heat-treatment and high dose irradiation

The aim of this paper is to report the sensitization of the TL peak appearing at 270 degrees C in the glow curve of natural quartz by using the combined effect of heat-treatments and irradiation with high gamma doses. For this, thirty discs with 6 x 1 mm(2) were prepared from plates parallell to a rhombolledral crystal face. The specimens were separated into four lots according to its TL read out between 160 and 320 degrees C. One lot was submitted to gamma doses of Co-60 radiation starting at 2 kGy and going up until a cumulative dose of 25 kGy. The other three lots were initially heal-treated at 500, 800 and 1000 degrees C and then irradiated with a single dose of 25kGy. The TL response of each lot was determined as a function of test-doses ranging from 0.1 to 30 mGy. As a result, it was observed that heat-treatments themselves did not produce the strong peak at 270 degrees C that was observed after the administration of high gamma doses. This peak is associated with the optical absorption band appearing at 470 rim which is due to the formation of [AlO4]degrees acting as electron-hole recombination centers. The formation of the 270 degrees C peak was preliminary analyzed in relation to aluminum- and oxygen-vacancy-related centers found in crystalline quartz. (C) 2008 Elsevier Ltd. All rights reserved.

Mechanisms of TL for production of the 230 degrees C peak in natural sodalite

The thermoluminescence (TL) peak in natural sodalite near 230 degrees C which appears only after submitted to thermal treatments and to gamma irradiation has been studied in parallel with electron paramagnetic resonance (EPR) spectrum appearing under the same procedure This study revealed a full correlation between the 230 degrees C TL peak and the eleven hyperfine lines from EPR spectrum In both case the centers disappear at the same temperature and are restored after gamma irradiation A complete model for the 230 C TL peak is presented and discussed In addition to the correlation and TL model specific characteristics of the TL peaks are described (C) 2010 Elsevier B V All rights reserved

Correlation between electron paramagnetic resonance and thermoluminescence in natural sodalite

Samples of natural sodalite, Na(8)Al(6)Si(6)O(24)Cl(2), submitted to gamma irradiation and to thermal treatments, have been investigated using the thermoluminescence (TL) and electron paramagnetic resonance (EPR) techniques. Both, natural and heat-treated samples at 500A degrees C in air for 30 min, present an EPR signal around g = 2.01132 attributed to oxygen hole centers. The EPR spectra of irradiated samples show an intense line at g = 2.0008 superimposed by a hyperfine multiplet of 11 lines due to an O(-) ion in an intermediate position with respect to two adjacent Al nuclei. In the TL measurements, the samples were annealed at 500A degrees C for 30 min and then irradiated with gamma doses varying from 0.001 to 20 kGy. All the samples have shown TL peaks at 110, 230, 270, 365, and 445A degrees C. A correlation between the EPR g = 2.01132 line and the 365A degrees C TL peak was observed. A TL model is proposed in which a Na(+) ion acts as a charge compensator when an Al(3+) ion replaces a Si(4+) lattice ion. The gamma ray destruction of the Al-Na complex provides an electron trapped at the Na and a hole trapped at a non-bridging oxygen ion adjacent to the Al(3+) ion.

Footemineite, the Mn-analog of atencioite, from the Foote mine, Kings Mountain, Cleveland County, North Carolina, USA, and its relationship with other roscherite-group minerals

Footemineite, ideally Ca2Mn2+square Mn22+Be4(PO4)(6)(OH)(4)-6H(2)O, triclinic, is a new member of the roscherite group. It occurs on thin fractures crossing quartz-microcline-spodumene pegmatite at the Foote mine, Kings Mountain, Cleveland County, North Carolina, U.S.A. Associated minerals are albite, analcime, eosphorite, siderite/rhodochrosite, fairfieldite, fluorapatite, quartz, milarite, and pyrite. Footemineite forms prismatic to bladed generally rough to barrel-shaped crystals up to about 1.5 mm long and I mm in diameter. Its color is yellow, the streak is white, and the luster is vitreous to slightly pearly. Footemineite is transparent and non-fluorescent. Twinning is simple, by reflection, with twin boundaries across the length of the crystals. Cleavage is good on {0 (1) over bar1}) and {100}. Density (calc.) is 2.873 g/cm(3). Footemineite is biaxial (-), n(alpha) = 1.620(2), n(beta) = 1.627(2), n(gamma) = 1.634(2) (white light). 2V(obs) = 80 degrees, 2V(calc) = 89.6 degrees. Orientation: X boolean AND b similar to 12 degrees, Y boolean AND c similar to 15 degrees, Z boolean AND a similar to 15 degrees. Elongation direction is c, dispersion: r > v or r < v, weak. Pleochroism: beta (brownish yellow) > alpha = gamma (yellow). Mossbauer and IR spectra are given. The chemical composition is (EDS mode electron microprobe, Li and Be by ICP-OES, Fe3+:Fe2+ y Mossbauer, H2O by TG data, wt%): Li2O 0.23, BeO 9.54, CaO 9.43, SrO 0.23, BaO 0.24, MgO 0.18, MnO 26.16, FeO 2.77, Fe2O3 0.62, Al2O3 0.14, P2O5 36.58, SiO2 0.42, H2O 13.1, total 99.64. The empirical formula is (Ca1.89Sr0.03Ba0.02)Sigma(1.94)(Mn-0.90(2+)square(0.10))Sigma(1.00)(square 0.78Li0.17Mg0.05) Sigma(1.00)(Mn3.252+Fe0.432+ Fe0.093+Al0.03)Sigma(3.80) Be-4.30(P5.81Si0.08O24)[(OH)3.64(H2O)0.36]Sigma(4.00)center dot 6.00H(2)O . The strongest reflection peaks of the powder diffraction pattern [d, angstrom (1, %) (hkl)] are 9.575 (53) (010), 5.998 (100) (0 (1) over bar1), 4.848 (26) (021), 3.192 (44) (210), 3.003 (14) (0 (2) over bar2), 2.803 (38) ((1) over bar 03), 2.650 (29) ((2) over bar 02), 2.424 (14) (231). Single-crystal unit-cell parameters are a = 6.788(2), b = 9.972(3), c = 10.014(2) A, (x = 73.84(2), beta = 85.34(2), gamma = 87.44(2)degrees,V = 648.74 angstrom(3), Z = 1. The space group is P (1) over bar. Crystal structure was refined to R = 0.0347 with 1273 independent reflections (F > 2(5). Footemineite is dimorphous with roscherite, and isostructural with atencioite. It is identical with the mineral from Foote mine described as ""triclinic roscherite."" The name is for the Foote mine, type locality for this and several other minerals.

All solid-state electrochromic devices with gelatin-based electrolyte

6 x 8cm(2) electrochromic devices (ECDs) with the configuration K-glass/EC-layer/electrotype/ion-storage (IS) layer/K-glass, have been assembled using Nb2O5:Mo EC layers, a (CeO2)(0.81)-TiO2 IS-layer and a new gelatin electrolyte containing Li+ ions. The structure of the electrolyte is X-ray amorphous. Its ionic conductivity passed by a maximum of 1.5 x 10(-5) S/CM for a lithium concentration of 0.3g/15ml. The value increases with temperature and follows an Arrhenius law with an activation energy of 49.5 kJ/mol. All solid-state devices show a reversible gray coloration, a long-term stability of more than 25,000 switching cycles (+/- 2.0 V/90 s), a transmission change at 550 nm between 60% (bleached state) and 40% (colored state) corresponding to a change of the optical density (Delta OD = 0. 15) with a coloration efficiency increasing from 10cm(2)/C (initial cycle) to 23cm(2)/C (25,000th cycle). (c) 2007 Elsevier B.V. All rights reserved.

Estudo do efeito inibidor do solicato de sódio na corrosão de uma liga de ali-mg-si, em meio de diferentes concentrações de cloreto

No presente trabalho, estudou-se o efeito inibidor do silicato de sódio na corrosão de uma liga de AI-Mg-Si em meio arejado e desarejado, contendo quantidades variadas de Ion cloreto no pW = 10,O . Nos diferentes tipos de ensaios realizados, tais como, traçado de curvas de polarização, ensaios galvanostáticos e com par galvânico, utilizou-se corpos de prova anodizados ou polidos. Os resultados experimentais mostraram que 1 g/l de silicato de sódio neutro de composição Na2O : 3.3 SiO2, demonstrou total efeito inibidor da corrosão da liga de A1-Mg-Si, em solução contendo até 60 p.p.m. de NaC1. Em concentrações de 1060 p.p,m. e maiores de NaC1, o silicato mostrou apenas efeito retardador da corrosão, com diminuição da incidência de pites. A análise dos produtos de corrosão, formadas sobre a superficie dos pites, feita usando-se raio-X e espectroscopia de infravermelho, mostrou que esses produtos têm estrutura amorfa e contêm silicato e grupos hidroxila. Finalmente, medidas de capacitância da dupla camada, confirmaram a existência de uma película que se forma sobre a superfície da liga de alumínio, quando em solução de silicato.

Influência de bolhas de hélio e da microestrutura sobre a evolução térmica de filmes de alumínio implantados com cobre

Este trabalho apresenta os resultados de um estudo sistemático sobre as influências do tamanho de grão de filmes finos de Al e da implantação de íons de He sobre a evolução térmica de distribuições de átomos de Cu e formação de precipitados de Al-Cu. Filmes finos de Al depositados sobre substrato de SiO2/Si através de dois processos diferentes foram implantados com íons de Cu+ e He+ produzindo uma solução sólida supersaturada de Al-Cu (≈ 2,5 a 3,5 at. %) e nano-bolhas de He. Os valores de energia dos íons foram escolhidos de tal forma a produzirem uma camada rica em Cu e He na região a aproximadamente 100nm da superfície. Tais filmes foram tratados termicamente em alto-vácuo nas temperaturas de 200ºC e 280ºC por tempos de 0,5h e 2h. Os filmes foram analisados por Retroespalhamento Rutherford, para determinação do perfil de concentração dos átomos de Cu, e por Microscopia Eletrônica de Transmissão, para determinação da microestrutura do Al e dos sistemas de nano-partículas Al-Cu e Al-He. Os resultados experimentais mostraram que a evolução térmica da distribuição dos átomos de Cu e a formação de precipitados de Al-Cu são significativamente afetadas pela configuração e tamanho de grão do filme de Al e pelas implantações de He. O presente estudo mostrou que existe uma forte correlação entre o fluxo de vacâncias e a estabilidade da microestrutura de filmes finos de Al (Al/SiO2/Si) implantados com íons de Cu+ e He+ e tratados termicamente. A possibilidade de controlar os fluxos de vacâncias através de configurações da microestrutura dos filmes de Al é, portanto, um tema de grande interesse tecnológico relacionado a durabilidade das interconexões metálicas de dispositivos microeletrônicos.