• Title/Summary/Keyword: cell sorting

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Micro-imaging techniques for evaluation of plastic microfluidic chip

  • Kim, Jung-Kyung;Hyunwoo Bang;Lee, Yongku;Chanil Chung;Yoo, Jung-Yul;Yang, Sang-Sik;Kim, Jin-Seung;Park, Sekwang;Chang, Jun-Keun
    • JSTS:Journal of Semiconductor Technology and Science
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    • v.1 no.4
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    • pp.239-247
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    • 2001
  • The Fluorescence-Activated Cell Sorter (FACS) is a well-established instrument used for identifying, enumerating, classifying and sorting cells by their physical and optical characteristics. For a miniaturized FACS device, a disposable plastic microchip has been developed which has a hydrodynamic focusing chamber using soft lithography. As the characteristics of the spatially confined sample stream have an effect on sample throughput, detection efficiency, and the accuracy of cell sorting, systematic fluid dynamic studies are required. Flow visualization is conducted with a laser scanning confocal microscopy (LSCM), and three-dimensional flow structure of the focused sample stream is reconstructed from 2D slices acquired at $1\mutextrm{m}$ intervals in depth. It was observed that the flow structure in the focusing chamber is skewed by unsymmetrical velocity profile arising from trapezoidal cross section of the microchannel. For a quantitative analysis of a microscopic flow structure, Confocal Micro-PIV system has been developed to evaluate the accelerated flow field in the focusing chamber. This study proposes a method which defines the depth of the measurement volume using a detection pinhole. The trajectories of red blood cells (RBCs) and their interactions with surrounding flow field in the squeezed sample stream are evaluated to find optimal shape of the focusing chamber and fluid manipulation scheme for stable cell transporting, efficient detection, and sorting

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Waveform Sorting of Rabbit Retinal Ganglion Cell Activity Recorded with Multielectrode Array (다채널전극으로 기록한 토끼 망막신경절세포의 활동전위 파형 구분)

  • Jin Gye Hwan;Lee Tae Soo;Goo Yang Sook
    • Progress in Medical Physics
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    • v.16 no.3
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    • pp.148-154
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    • 2005
  • Since the output of retina for visual stimulus is carried by neurons of very diverse functional properties, it is not adequate to use conventional single electrode for recording the retinal action potential. For this purpose, we used newly developed multichannel recording system for monitoring the simultaneous electrical activities of many neurons in a functioning piece of retina. Retinal action potentials are recorded with an extra-cellular planar array of 60 microelectrodes. In studying the collective activity of the ganglion cell population it is essential to recognize basic functional distinctions between individual neurons. Therefore, it is necessary to detect and to classify the action potential of each ganglion cell out of mixed signal. We programmed M-files with MATLAB for this sorting process. This processing is mandatory for further analysis, e.g. poststimulus time histogram (PSTH), auto-correlogram, and cross-correlogram. We established MATLAB based protocol for waveform classification and verified that this approach was effective as an initial spike sorting method.

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Rapid bacterial identification using Raman spectroscopy (라만 분광법을 활용한 세균 검측 기술)

  • No, Jee Hyun;Lee, Tae Kwon
    • Korean Journal of Microbiology
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    • v.53 no.2
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    • pp.71-78
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    • 2017
  • Raman microspectroscopy is a promising tool for microbial analysis at single cell level since it can rapidly measure the cell materials including lipids, nucleic acids, and proteins by measuring the inelastic scattering of a molecule irradiated by monochromatic lights. Using Raman spectra provides high specificity and sensitivity in classification of bacteria at the strain level. In addition, a Raman approach coupled with stabled isotope such as $^{13}C$ and $^2H$ is able to detect and quantify general metabolic activity at single cell level. After bacterial detection process by Raman microspectroscopy, interested unculturable cell sorting and single cell genomics can be accomplished by combination with optical tweezer and microfluidic devices. In this review, the characteristics and applications of Raman microspectroscopy were reviewed and summarized in order to provide a better understanding of microbial analysis using Raman spectroscopy.

Purification of Pig Muscle Stem Cells Using Magnetic-Activated Cell Sorting (MACS) Based on the Expression of Cluster of Differentiation 29 (CD29)

  • Choi, Kwang-Hwan;Kim, Minsu;Yoon, Ji Won;Jeong, Jinsol;Ryu, Minkyung;Jo, Cheorun;Lee, Chang-Kyu
    • Food Science of Animal Resources
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    • v.40 no.5
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    • pp.852-859
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    • 2020
  • The muscle stem cells of domestic animals are of interest to researchers in the food and biotechnology industries for the production of cultured meat. For producing cultured meat, it is crucial for muscle stem cells to be efficiently isolated and stably maintained in vitro on a large scale. In the present study, we aimed to optimize the method for the enrichment of pig muscle stem cells using a magnetic-activated cell sorting (MACS) system. Pig muscle stem cells were collected from the biceps femoris muscles of 14 d-old pigs of three breeds [Landrace×Yorkshire×Duroc (LYD), Berkshire, and Korean native pigs] and cultured in skeletal muscle growth medium-2 (SkGM-2) supplemented with epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580). Approximately 30% of total cultured cells were nonmyogenic cells in the absence of purification in our system, as determined by immunostaining for cluster of differentiation 56 (CD56) and CD29, which are known markers of muscle stem cells. Interestingly, following MACS isolation using the CD29 antibody, the proportion of CD56+/CD29+ muscle stem cells was significantly increased (91.5±2.40%), and the proportion of CD56 single-positive nonmyogenic cells was dramatically decreased. Furthermore, we verified that this method worked well for purifying muscle stem cells in the three pig breeds. Accordingly, we found that CD29 is a valuable candidate among the various marker genes for the isolation of pig muscle stem cells and developed a simple sorting method based on a single antibody to this protein.

Advanced OS-CFAR Processor Design with Low Computational Effort (순서통계에 근거한 개선된 CFAR 검파기의 하드웨어 구조 제안)

  • Hyun, Eu-Gin;Lee, Jong-Hun
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.16 no.1
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    • pp.65-71
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    • 2012
  • An OS-CFAR (Ordered Statistics CFAR) based on a sorting algorithm is useful for automotive radar systems in a multi-target situation. However, while the typical cell-averaging CFAR has low computational complexity, the OS-CFAR has much higher computation effort. In this paper, we design the new OS-CFAR architecture with a low computational effort. In the proposed method, since one time sorting processing is performed for the decision of the CFAR threshold, the whole processing effort can be reduced. When the fast sorting technique is employed, the computing time of the proposed OS-CFAR is always much shorter compared with typical OS-CFAR method regardless of the data size. We also present the processing result of proposed architecture using the real radar data.

Establishment of B-1 cell-derived polyreactive monoclonal antibodies and expression of costimulators by B-cell to antigenic stimulation (B-1 세포 유래 다중반응성 단클론 항체의 형성과 항원 자극에 대한 B 세포의 동시자극자 발현)

  • Lee, Ju-Youn;Choi, Jeom-Il;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.371-384
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    • 2007
  • 연구 목적 : 면역 반응에서 B-1 세포의 정확한 역할은 아직 명확히 규명되지 않았으나 최근 B-1 세포가 면역의 내성을 야기하고 유지하는데 필요한 특성들을 가지고 있음이 밝혀지고 있다. 이에 본 연구에서는 B-1 세포 유래 단클론 항체의 특성과 항원 자극에 대한 B 세포의 동시자극자 (MHC Class, B7-2, 7-2) 발현을 평가하여 B 세포의 면역조절 기능을 알아보고자 한다. 연구 대상 및 방법 : B-1 세포 유래 단클론 항체를 형성하는 잡종세포주를 이용하여 다양한 내, 외인성 항원에 대한 단클론 항체의 반응 양상을 평가하였다. 여러 내, 외인성 항원으로 면역한 쥐의 복강과 비장 B 세포의 동시자극자의 발현을 Fluorescence Activated Cell Sorting (FACS)을 이용하여 평가하였다. 결과 : 최종적으로 단클론을 형성하는 2개의 클론을 형성하였고, 이 B-1 세포 유래 단클론 항체는 dose-saturable pattern을 띄는 다중 반응성을 나타내었다. FACS를 이용한 동시자극자의 발현 검사에서는 MHC 발현은 복강과 비장의 B 세포가 유사하였으나, B7-1과 7-2는 복강의 B 세포에서 더 뚜렷한 발현을 보여주었다. 결론 : B-1 세포 유래 단클론 항체는 다양한 내, 외인성 항원 자극에 대해 dose-saturable한 다중반응성을 나타낸다. 복강과 비장의 B세포는 내, 외인성 항원의 면역에 있어서 동시자극자 발현이 명확히 다른 양상을 나타냄을 확인할 수 있었다.

Development of Rapid Antibody-based Therapeutic Platform Correspondence for New Viruses Using Antigen-specific Single Cell Memory B Cell Sorting Technology (항원 특이적 단일 기억 B 세포 분리를 이용한 신종 바이러스 대응 신속 항체 플랫폼 개발)

  • Jiyoon Seok;Suhan Jung;Ye Gi Han;Arum Park;Jung Eun Kim;Young Jo Song;Chi Ho Yu;Hyeongseok Yun;Se Hun Gu;Seung-Ho Lee;Yong Han Lee;Gyeunghaeng Hur;Woong Choi
    • Journal of the Korea Institute of Military Science and Technology
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    • v.27 no.1
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    • pp.116-125
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    • 2024
  • The COVID-19 pandemic is not over despite the emergency use authorization as can see recent COVID-19 daily confirmed cases. The viruses are not only difficult to diagnose and treat due to random mutations, but also pose threat human being because they have the potential to be exploited as biochemical weapons by genetic manipulation. Therefore, it is inevitable to the rapid antibody-based therapeutic platform to quickly respond to future pandemics by new/re-emerging viruses. Although numerous researches have been conducted for the fast development of antibody-based therapeutics, it is sometimes hard to respond rapidly to new viruses because of complicated expression or purification processes for antibody production. In this study, a novel rapid antibody-based therapeutic platform using single B cell sorting method and mRNA-antibody. High immunogenicity was caused to produce antibodies in vivo through mRNA-antigen inoculation. Subsequently, antigen-specific antibody candidates were selected and obtained using isolation of B cells containing antibody at the single cell level. Using the antibody-based therapeutic platform system in this study, it was confirmed that novel antigen-specific antibodies could be obtained in about 40 days, and suggested that the possibility of rapid response to new variant viruses.

Performance Analysis of Output Queued Batcher-Banyan Switch for ATM Network (ATM 망에 적용 가능한 출력단 버퍼형 Batcher-Banyan 스위치의 성능분석)

  • Keol-Woo Yu;Kyou Ho Lee
    • Journal of the Korea Society for Simulation
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    • v.8 no.4
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    • pp.1-8
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    • 1999
  • This paper proposes an ATM switch architecture called Output Queued Batcher-Banyan switch (OQBBS). It consists of a Sorting Module, Expanding Module, and Output Queueing Modules. The principles of channel grouping and output queueing are used to increase the maximum throughput of an ATM switch. One distinctive feature of the OQBBS is that multiple cells can be simultaneously delivered to their desired output. The switch architecture is shown to be modular and easily expandable. The performance of the OQBBS in terms of throughput, cell delays, and cell loss rate under uniform random traffic condition is evaluated by computer simulation. The throughput and the average cell delay are close to the ideal performance behavior of a fully connected output queued crossbar switch. It is also shown that the OQBBS meets the cell loss probability requirement of $10^{-6}$.

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Optimization of Yeast Surface-Displayed cDNA Library Screening for Low Abundance Targets

  • Kim, Juhyung;Kim, Hyung Kyu;Jang, Hye Jeong;Kim, Eunkyung;Kim, Moon Kyu
    • Journal of Microbiology and Biotechnology
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    • v.25 no.4
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    • pp.547-553
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    • 2015
  • The yeast surface-displayed cDNA library has been used to identify unknown antigens. However, when unknown target antigens show moderate-to-low abundance, some modifications are needed in the screening process. In this study, a directional random-primed cDNA library was used to increase the number of candidates for the unknown antigen. To avoid the loss of target yeast clones that express proteins at a low frequency in the cDNA library, a comprehensive monitoring system based on magnetic-activated cell sorting, fluorescence-activated cell sorting, and immunofluorescence was established, and a small number of target yeast cells was successfully enriched. These results showed that our optimized method has potential application for identifying rare unknown antigens of the human monoclonal antibody.

Single Buffer types of ATM Switches based on Circulated Priority Algorithm (순환적 순위 알고리즘을 이용한 단일형 버퍼형태의 ATM스위치)

  • Park Byoung-soo;Cho Tae-kyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.5 no.5
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    • pp.429-432
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    • 2004
  • In this paper, we propose a new sorting algorithm for ATM switch with a shared buffer which has a sequencer architecture with single queue. The proposed switch performs a sorting procedure of ATM cell based on the output port number of ATM cell with hardware implementation. The proposed architecture has a single buffer physically but logically it has function of multi-queue which is designed at most to control the conflicts in output port. In the future, this architecture will take various applications for routing switch and has flexibility for the extension of system structure. therefore, this structure is expected on good structure in effective transmission.

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