• Title/Summary/Keyword: Cell micropatterns

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Chemical Imaging Analysis of the Micropatterns of Proteins and Cells Using Cluster Ion Beam-based Time-of-Flight Secondary Ion Mass Spectrometry and Principal Component Analysis

  • Shon, Hyun Kyong;Son, Jin Gyeong;Lee, Kyung-Bok;Kim, Jinmo;Kim, Myung Soo;Choi, Insung S.;Lee, Tae Geol
    • Bulletin of the Korean Chemical Society
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    • v.34 no.3
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    • pp.815-819
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    • 2013
  • Micropatterns of streptavidin and human epidermal carcinoma A431 cells were successfully imaged, as received and without any labeling, using cluster $Au_3{^+}$ ion beam-based time-of-flight secondary ion mass spectrometry (TOF-SIMS) together with a principal component analysis (PCA). Three different analysis ion beams ($Ga^+$, $Au^+$ and $Au_3{^+}$) were compared to obtain label-free TOF-SIMS chemical images of micropatterns of streptavidin, which were subsequently used for generating cell patterns. The image of the total positive ions obtained by the $Au_3{^+}$ primary ion beam corresponded to the actual image of micropatterns of streptavidin, whereas the total positive-ion images by $Ga^+$ or $Au^+$ primary ion beams did not. A PCA of the TOF-SIMS spectra was initially performed to identify characteristic secondary ions of streptavidin. Chemical images of each characteristic ion were reconstructed from the raw data and used in the second PCA run, which resulted in a contrasted - and corrected - image of the micropatterns of streptavidin by the $Ga^+$ and $Au^+$ ion beams. The findings herein suggest that using cluster-ion analysis beams and multivariate data analysis for TOF-SIMS chemical imaging would be an effectual method for producing label-free chemical images of micropatterns of biomolecules, including proteins and cells.

Fabrication of PDMS Stencil using Gas Blowing for Micropatterned 3T3 Cell Culture (가스 블로잉을 이용한 PDMS 스텐실 제작 및 3T3 세포의 마이크로 패터닝)

  • Choi, Jin Ho;Kim, Gyu Man
    • Journal of the Korean Society for Precision Engineering
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    • v.30 no.2
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    • pp.236-240
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    • 2013
  • In this presentation, we propose a fabrication method of PDMS stencil to apply into a localized culture of NIH/3T3 cells. PDMS stencil was fabricated by nitrogen gas blowing and soft lithography from preparing SU-8 master mold by photolithography. PDMS stencil pattern was production of the circle size 20 to $500{\mu}m$. In the culture test of PDMS stencil, a stencil was placed on a glass disk. The NIH/3T3 cells were successfully cultured into micropatterns by using the PDMS stencil. The results showed that cells could be cultured into micropatterns with precisely controlled manner at any shapes and specific size for bioscience study and bioengineering applications.

Quantitative Analysis of Growth of Cells on Physicochemically Modified Surfaces

  • Chandra, Prakash;Kim, Jihee;Rhee, Seog Woo
    • Bulletin of the Korean Chemical Society
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    • v.34 no.2
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    • pp.524-530
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    • 2013
  • In this study, we describe the most expected behavior of cells on the modified surface and the correlation between the modified substrates and the response of cells. The physicochemical characteristics of substrates played an essential role in the adhesion and proliferation of cells. Glass and polymer substrates were modified using air plasma oxidation, and the surfaces were coated with self-assembled monolayer molecules of silanes. The PDMS substrates embedded with parallel micropatterns were used for evaluation of the effect of topologically modified substrate on cellular behaviour. BALB/3T3 fibroblast cells were cultured on different surfaces with distinct wettability and topology, and the growth rates and morphological change of cells were analyzed. Finally, we found the optimum conditions for the adhesion and proliferation of cells on the modified surface. This study will provide insight into the cell-surface interaction and contribute to tissue engineering applications.

Simple Formation of Poly(sodium 4-styrenesulfonate) Pattern on the Hydrophobic Substrate for the Control of Cell Adhesion via a Selective Ion Irradiation

  • Kim, Soo-Jung;Hwang, In-Tae;Jung, Jin-Mook;Jung, Chan-Hee
    • Journal of Radiation Industry
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    • v.7 no.2_3
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    • pp.149-154
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    • 2013
  • In this study, the simple preparation of poly(sodium 4-styrenesulfonate) (PSS)-patterned substrate via a selective ion irradiation was investigated to manipulate cell adhesion. PSS thin films spin-coated onto the hydrophobic polystyrene (PS) was patterned through masked 150 keV proton irradiation followed by developing with deionized water. The characteristics of the resulting PSS-patterned surfaces were investigated by using microscope, surface profiler, FT-IR, XPS, and contact angle analyzer. These analytical results revealed that the resolved $100{\mu}m$ PSS patterns were formed on the hydrophobic PS surface above the fluence of $1{\times}10^{15}ions\;cm^{-2}$ and the chemical structure, composition, and wettability of the PSS patterns were dependant on a fluence. Moreover, the results of the in-vitro cell culture and proliferation assay exhibited that H1299 cells preferentially adhered and proliferated onto the more hydrophilic PSS part of the PSS-patterned PS and the well-aligned cell patterns was formed on the PSS-patterned PS particularly at the fluence of $1{\times}10^{15}ions\;cm^{-2}$.

Constructing a Three-Dimensional Endothelial Cell Layer in a Circular PDMS Microchannel

  • Choi, Jong Seob;Piao, Yunxian;Kim, Kyung Hoon;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.274.2-274.2
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    • 2013
  • We described a simple and efficient fabrication method for generating microfluidic channels with a circular-cross sectional geometry by exploiting the reflow phenomenon of a thick positive photoresist. Initial rectangular shaped positive photoresist micropatterns on a silicon wafer, which were fabricated by a conventional photolithography process, were converted into a half-circular shape by tuning the temperature to around $105^{\circ}C$. Through optimization of the reflow conditions, we could obtain a perfect circular micropattern of the positive photoresist, and control the diameter in a range from 100 to 400 ${\mu}m$. The resultant convex half-circular photoresist was used as a template for fabricating a concave polydimethylsiloxane (PDMS) through a replica molding process, and a circular PDMS microchannel was produced by bonding two half-circular PDMS layers. A variety of channel dimensions and patterns can be easily prepared, including straight, S-curve, X-, Y-, and T-shapes to mimic an in vivo vascular network. To inform an endothelial cell layer, we cultured primary human umbilical vein endothelial cells (HUVECs) inside circular PDMS microchannels, and demonstrated successful cell adhesion, proliferation, and alignment along the channel.

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Fabrication of Biodegradable Disc-shaped Microparticles with Micropattern using a Hot Embossing Process with Porous Microparticles

  • Hwang, Ji-Yea;Choy, Young-Bin;Seo, Soon-Min;Park, Jung-Hwan
    • Journal of Pharmaceutical Investigation
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    • v.41 no.3
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    • pp.147-151
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    • 2011
  • This paper demonstrates the development of a method for preparing micropatterned microdiscs in order to increase contact area with cells and to change the release pattern of drugs. The microdiscs were manufactured with hot embossing, where a polyurethane master structure was pressed onto both solid and porous microparticles made of polylactic-co-glycolic acid at various temperatures to form a micropattern on the microdiscs. Flat microdiscs were formed by hot embossing of porous microparticles; the porosity allowed space for flattening of the microdiscs. Three types of micro-grooves were patterned onto the flat microdiscs using prepared micropatterned molds: (1) 10 ${\mu}M$ deep, 5 ${\mu}M$ wide, and spaced 2 ${\mu}M$ apart; (2) 10 ${\mu}M$ deep, 9 ${\mu}M$ wide, and spaced 5 ${\mu}M$ apart; and (3) 10 ${\mu}M$ deep, 50 ${\mu}M$ wide, and spaced 50 ${\mu}M$ apart. This novel microdisc preparation method using hot embossing to create micropatterns on flattened porous microparticles provides the opportunity for low-cost, rapid manufacture of microdiscs that can be used to control cell adhesion and drug delivery rates.