• Title/Summary/Keyword: Glass coverslip

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TEM Sample Preparation for Cultured Neurons on a Glass Coverslip (Hydrofluoric acid 용액을 이용한 유리 커버글라스에 배양된 신경세포의 전자현미경 시료제작법)

  • Oh, Hyun-Woo;Park, Ho-Yong
    • Applied Microscopy
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    • v.35 no.4
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    • pp.10-15
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    • 2005
  • Cultured neurons from Drosophila brain on a glass coverslip to understand the structural basis of synapse were prepared for TEM observations. Neurons on a coverslip were fixed, dehydrated and embedded in Epon without separating from coverslip. After polymerization, the block was placed in 49% hydrofluoric acid to remove the coverslip. The block was examined under a light microscope to select exact neurons, then trimmed and sectioned for TEM observation.

Growth and Migration of BALB/3T3 Fibroblast Cells on Nano-engineered Silica Beads Surface

  • Kim, Jihee;Chandra, Prakash;Yang, Jiyoon;Rhee, Seog Woo
    • Bulletin of the Korean Chemical Society
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    • v.34 no.12
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    • pp.3715-3721
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    • 2013
  • In this study, the behavior of cells on the modified surface, and the correlation between the modified substrates and the response of cells is described. A close-packed layer of nano-sized silica beads was prepared on a coverslip, and the adhesion, proliferation, and migration of BALB/3T3 fibroblast cells on the silica layer was monitered. The 550 nm silica beads were synthesized by the hydrolysis and condensation reaction of tetraethylorthosilicate in basic solution. The amine groups were introduced onto the surfaces of silica particles by treatment with 3-aminopropyltrimethoxysilane. The close-packed layer of silica beads on the coverslip was obtained by the reaction of the amine-functionalized silica beads and the (3-triethoxysilyl)propylsuccinic anhydride treated coverslip. BALB/3T3 fibroblast cells were loaded on bare glass, APTMS coated glass, and silica bead coated glass with the same initial cell density, and the migration and proliferation of cells on the substrates was investigated. The cells were fixed and stained with antibodies in order to analyze the changes in the actin filaments and nuclei after culture on the different surfaces. The motility of cells on the silica bead coated glass was greater than that of the cells cultured on the control substrate. The growth rate of cells on the silica bead coated glass was slower than that of the control. Because the close-packed layer of silica beads gave an embossed surface, the adhesion of cells was very weak compared to the smooth surfaces. These results indicate that the adhesion of cells on the substrates is very important, and the actin filaments might play key roles in the migration and proliferation of cells. The nuclei of the cells were shrunk on the weakly adhered surfaces, and the S1 stage in which DNA is duplicated in the cell dividing processes might be retarded. As a result, the rate of proliferation of cells was decreased compared to the smooth surface of the control. In conclusion, the results described here are very important in the understanding of the interaction between implanted materials and biosystems.

Adhesion and Biofilm Formation Abilities of Bacteria Isolated from Dental Unit Waterlines (치과용 유니트 수관에서 분리한 세균의 부착 및 바이오필름 형성 능력)

  • Yoon, Hye Young;Lee, Si Young
    • Journal of dental hygiene science
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    • v.18 no.2
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    • pp.69-75
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    • 2018
  • The purpose of our study is to compare the adhesion and biofilm formation abilities of isolates from water discharged from dental unit waterlines (DUWLs). Bacteria were isolated from a total of 15 DUWLs. Twelve isolates were selected for the experiment. To confirm the adhesion ability of the isolates, each isolate was attached to a glass coverslip using a 12-well plate. Plates were incubated at $26^{\circ}C$ for 7 days, and the degree of adhesion of each isolate was scored. To verify the biofilm formation ability of each isolate, biofilms were allowed to form on a 96-well polystyrene flat-bottom microtiter plate. The biofilm accumulations of all isolates formed at $26^{\circ}C$ for 7 days were identified and compared. A total of 56 strains were isolated from 15 water samples including 12 genera and 31 species. Of the 56 isolates, 12 isolates were selected according to the genus and used in the experiment. Sphingomonas echinoides, Methylobacterium aquaticum, and Cupriavidus pauculus had the highest adhesion ability scores of +3 among 12 isolates. Among these three isolates, the biofilm accumulation of C. pauculus was the highest and that of S. echinoides was the third-most abundant. The lowest biofilm accumulations were identified in Microbacterium testaceum and M. aquaticum. Most isolates with high adhesion ability also exhibited high biofilm formation ability. Analysis of adhesion and biofilm formation of the isolates from DUWLs can provide useful information to understand the mechanism of DUWL biofilm formation and development.

Cell-cultivable ultrasonic transducer integrated on glass-coverslip (세포 배양 가능한 커버슬립형 초음파 변환자)

  • Keunhyung Lee;Jinhyoung Park
    • The Journal of the Acoustical Society of Korea
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    • v.42 no.5
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    • pp.412-421
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    • 2023
  • Ultrasound brain stimulation is spot-lighted by its capability of inducing brain cell activation in a localized deep brain region and ultimately treating impaired brain function while the efficiency and directivity of neural modulation are highly dependent on types of stimulus waveforms. Therefore, to optimize the types of stimulation parameters, we propose a cell-cultivable ultrasonic transducer having a series stack of a spin-coated polymer piezoelectric element (Poly-vinylidene fluoride-trifluorethylene, PVDF-TrFE) and a parylene insulating layer enhancing output acoustic pressure on a glass-coverslip which is commonly used in culturing cells. Due to the uniformity and high accuracy of stimulus waveform, tens of neuronal cell responses located on the transducer surface can be recorded simultaneously with fluorescence microscopy. By averaging the cell response traces from tens of cells, small changes to the low intensity ultrasound stimulations can be identified. In addition, the reduction of stimulus distortions made by standing wave generated from reflections between the transducers and other strong reflectors can be achieved by placing acoustic absorbers. Through the proposed ultrasound transducer, we could successfully observe the calcium responses induced by low-intensity ultrasound stimulation of 6 MHz, 0.2 MPa in astrocytes cultured on the transducer surface.

Quantitative Determination of 3D-Printing and Surface-Treatment Conditions for Direct-Printed Microfluidic Devices

  • Hyun Namgung;Abdi Mirgissa Kaba;Hyeonkyu Oh;Hyunjin Jeon;Jeonghwan Yoon;Haseul Lee;Dohyun Kim
    • BioChip Journal
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    • v.16
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    • pp.82-98
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    • 2020
  • We report a quantitative and systematic method for determining 3D-printing and surface-treatment conditions that can help improve the optical quality of direct-printed microfluidic devices. Digital light processing (DLP)-stereolithography (SLA) printing was extensively studied in microfluidics owing to the rapid, one-step, cleanroom-free, maskless, and high-definition microfabrication of 3D-microfluidic devices. However, optical imaging or detection for bioassays in DLP-SLA-printed microfluidic devices are limited by the translucence of photopolymerized resins. Various approaches, including mechanical abrasions, chemical etching, polymer coatings, and printing on transparent glass/plastic slides, were proposed to address this limitation. However, the effects of these methods have not been analyzed quantitatively or systematically. For the first time, we propose quantitative and methodological determination of 3D-printing and surface-treatment conditions, based on optical-resolution analysis using USAF 1951 resolution test targets and a fluorescence microbead slide through 3D-printed coverslip chips. The key printing parameters (resin type, build orientation, layer thickness, and layer offset) and surface-treatment parameters (grit number for sanding, polishing time with alumina slurry, and type of refractive-index-matching coatings) were determined in a step-wise manner. As a result, we achieved marked improvements in resolution (from 80.6 to 645.1 lp/mm) and contrast (from 3.30 to 27.63% for 645.1 lp/mm resolution). Furthermore, images of the fluorescence microbeads were qualitatively analyzed to evaluate the proposed 3D-printing and surface-treatment approach for fluorescence imaging applications. Finally, the proposed method was validated by fabricating an acoustic micromixer chip and fluorescently visualizing cavitation microstreaming that emanated from an oscillating bubble captured inside the chip. We expect that our approach for enhancing optical quality will be widely used in the rapid manufacturing of 3D-microfluidic chips for optical assays.