The Experts below are selected from a list of 312 Experts worldwide ranked by ideXlab platform

Jun Miao - One of the best experts on this subject based on the ideXlab platform.

  • rapid isolation of single malaria parasite infected red blood Cells by Cell sorting
    Nature Protocols, 2011
    Co-Authors: Jun Miao, Liwang Cui
    Abstract:

    Malaria research often requires isolation of individually infected red blood Cells (RBCs) or of a homogenous parasite population derived from a single parasite (clone). Traditionally, isolation of individual, parasitized RBCs or parasite cloning is achieved by limiting dilution or micromanipulation. This protocol describes a method for more efficient cloning of the malaria parasite; the method uses a Cell Sorter to rapidly isolate Plasmodium falciparum-infected RBCs singly. By gating the parameters of forward-angle light scatter and side-angle light scatter in a Cell Sorter, singly infected RBCs can be isolated and automatically deposited into a 96-well culture plate within 1 min. Including a Percoll purification step; the entire procedure to seed a 96-well plate with singly infected RBCs can take <40 min. This highly efficient single-Cell sorting protocol should be useful for cloning of both laboratory parasite populations from genetic manipulation experiments and clinical samples.

  • rapid isolation of single malaria parasite infected red blood Cells by Cell sorting
    Nature Protocols, 2011
    Co-Authors: Jun Miao
    Abstract:

    Malaria research often requires isolation of individually infected red blood Cells (RBCs) or a homogenous parasite population derived from a single parasite (clone). Traditionally, isolation of individual, parasitized RBCs or parasite cloning is achieved by limiting dilution or micromanipulation. This protocol describes a method for more efficient cloning of the malaria parasite, which uses a Cell Sorter to rapidly isolate singly Plasmodium falciparum-infected RBCs. By gating the parameters of forward angle light scatter (FSC) and side angle light scatter (SSC) in a Cell Sorter, singly-infected RBCs can be isolated and automatically deposited into a 96-well culture plate within one minute. To include a Percoll purification step, the entire procedure to seed a 96-well plate with singly-infected RBCs takes less than 40 min. This highly efficient single-Cell sorting protocol should be useful for cloning of both laboratory parasite populations from genetic manipulation experiments and clinical samples.

Pei-yu Chiou - One of the best experts on this subject based on the ideXlab platform.

  • Pulsed laser activated Cell Sorter with dielectrophoretic single stream sheathless focusing
    2017 IEEE 30th International Conference on Micro Electro Mechanical Systems (MEMS), 2017
    Co-Authors: Xiongfeng Zhu, Yu-chun Kung, Michael A. Teitell, Pei-yu Chiou
    Abstract:

    We report a novel sheathless microfluidic fluorescence-activated Cell Sorter utilizing size-independent three-dimensional dielectrophoretic (DEP) single stream focusing and pulsed laser activated Cell sorting (PLACS). This is realized by fabricating a 3D microfluidic device using two glass substrates with patterned electrodes sandwiching a thin and open PDMS channel. DEP forces are provided along a 4cm-long channel that focuses particles into a single stream passing through a fluorescence detection zone regardless of their sizes and types at high flow speeds. Pulsed laser-induced cavitation bubbles are used to provide rapid (∼30 μs) and precise fluid perturbation to sort out particles upon the detection of fluorescence. High purity sorting has been accomplished at a throughput of 1500 particles sec−1.

  • pulsed laser activated Cell Sorter placs for high throughput fluorescent mammalian Cell sorting
    Proceedings of SPIE, 2014
    Co-Authors: Yue Chen, Michael A. Teitell, Dino Di Carlo, Aram J Chung, Yu Chung Kung, Pei-yu Chiou
    Abstract:

    We present a Pulsed Laser Activated Cell Sorter (PLACS) realized by exciting laser induced cavitation bubbles in a PDMS microfluidic channel to create high speed liquid jets to deflect detected fluorescent samples for high speed sorting. Pulse laser triggered cavitation bubbles can expand in few microseconds and provide a pressure higher than tens of MPa for fluid perturbation near the focused spot. This ultrafast switching mechanism has a complete on-off cycle less than 20 μsec. Two approaches have been utilized to achieve 3D sample focusing in PLACS. One is relying on multilayer PDMS channels to provide 3D hydrodynamic sheath flows. It offers accurate timing control of fast (2 m sec-1) passing particles so that synchronization with laser bubble excitation is possible, an critically important factor for high purity and high throughput sorting. PLACS with 3D hydrodynamic focusing is capable of sorting at 11,000 Cells/sec with >95% purity, and 45,000 Cells/sec with 45% purity using a single channel in a single step. We have also demonstrated 3D focusing using inertial flows in PLACS. This sheathless focusing approach requires 10 times lower initial Cell concentration than that in sheath-based focusing and avoids severe sample dilution from high volume sheath flows. Inertia PLACS is capable of sorting at 10,000 particles sec-1 with >90% sort purity.

  • 3d pulsed laser triggered high speed microfluidic fluorescence activated Cell Sorter
    Analyst, 2013
    Co-Authors: Yue Chen, Yu-chun Kung, Michael A. Teitell, Pei-yu Chiou
    Abstract:

    We report a 3D microfluidic pulsed laser-triggered fluorescence-activated Cell Sorter capable of sorting at a throughput of 23 000 Cells per s with 90% purity in high-purity mode and at a throughput of 45 000 Cells per s with 45% purity in enrichment mode in one stage and in a single channel. This performance is realized by exciting laser-induced cavitation bubbles in a 3D PDMS microfluidic channel to generate high-speed liquid jets that deflect detected fluorescent Cells and particles focused by 3D sheath flows. The ultrafast switching mechanism (20 μs complete on–off cycle), small liquid jet perturbation volume, and three-dimensional sheath flow focusing for accurate timing control of fast (1.5 m s−1) passing Cells and particles are three critical factors enabling high-purity sorting at high-throughput in this Sorter.

  • Pulsed laser triggered high speed microfluidic fluorescence activated Cell Sorter
    2012 IEEE 25th International Conference on Micro Electro Mechanical Systems (MEMS), 2012
    Co-Authors: Yue Chen, Michael A. Teitell, Dino Di Carlo, Sung Yong Park, Jason S. Hong, Tara Teslaa, Jiang F. Zhong, Pei-yu Chiou
    Abstract:

    We report a high speed and high purity pulsed laser triggered fluorescence activated Cell Sorter (PLACS) with a sorting throughput up to 20,000 mammalian Cells/s with 37% sorting purity, 90% Cell viability in enrichment mode, and >90% purity in high purity mode at 1,500 Cells/s or 3,000 beads/s. Fast switching (30 μs) and a small perturbation volume (∼90 pL) is realized by a unique sorting mechanism in which explosive vapor bubbles are generated using focused laser pulses in a single layer microfluidic PDMS channel.

Liwang Cui - One of the best experts on this subject based on the ideXlab platform.

  • rapid isolation of single malaria parasite infected red blood Cells by Cell sorting
    Nature Protocols, 2011
    Co-Authors: Jun Miao, Liwang Cui
    Abstract:

    Malaria research often requires isolation of individually infected red blood Cells (RBCs) or of a homogenous parasite population derived from a single parasite (clone). Traditionally, isolation of individual, parasitized RBCs or parasite cloning is achieved by limiting dilution or micromanipulation. This protocol describes a method for more efficient cloning of the malaria parasite; the method uses a Cell Sorter to rapidly isolate Plasmodium falciparum-infected RBCs singly. By gating the parameters of forward-angle light scatter and side-angle light scatter in a Cell Sorter, singly infected RBCs can be isolated and automatically deposited into a 96-well culture plate within 1 min. Including a Percoll purification step; the entire procedure to seed a 96-well plate with singly infected RBCs can take <40 min. This highly efficient single-Cell sorting protocol should be useful for cloning of both laboratory parasite populations from genetic manipulation experiments and clinical samples.

Tony Jun Huang - One of the best experts on this subject based on the ideXlab platform.

  • standing surface acoustic wave ssaw based fluorescence activated Cell Sorter
    Small, 2018
    Co-Authors: Liqiang Ren, Yuchao Chen, Lin Wang, Zhangming Mao, Pohsun Huang, Shujie Yang, Peiran Zhang, Tony Jun Huang
    Abstract:

    Microfluidic fluorescence-activated Cell Sorters (μFACS) have attracted considerable interest because of their ability to identify and separate Cells in inexpensive and biosafe ways. Here a high-performance μFACS is presented by integrating a standing surface acoustic wave (SSAW)-based, 3D Cell-focusing unit, an in-plane fluorescent detection unit, and an SSAW-based Cell-deflection unit on a single chip. Without using sheath flow or precise flow rate control, the SSAW-based Cell-focusing technique can focus Cells into a single file at a designated position. The tight focusing of Cells enables an in-plane-integrated optical detection system to accurately distinguish individual Cells of interest. In the acoustic-based Cell-deflection unit, a focused interdigital transducer design is utilized to deflect Cells from the focused stream within a minimized area, resulting in a high-throughput sorting ability. Each unit is experimentally characterized, respectively, and the integrated SSAW-based FACS is used to sort mammalian Cells (HeLa) at different throughputs. A sorting purity of greater than 90% is achieved at a throughput of 2500 events s-1 . The SSAW-based FACS is efficient, fast, biosafe, biocompatible and has a small footprint, making it a competitive alternative to more expensive, bulkier traditional FACS.

  • a high throughput acoustic Cell Sorter
    Lab on a Chip, 2015
    Co-Authors: Liqiang Ren, Yuchao Chen, Lin Wang, Stewart J. Levine, Zhangming Mao, Pohsun Huang, Joseph Rufo, Feng Guo, Philip J Mccoy, Tony Jun Huang
    Abstract:

    Acoustic-based fluorescence activated Cell Sorters (FACS) have drawn increased attention in recent years due to their versatility, high biocompatibility, high controllability, and simple design. However, the sorting throughput for existing acoustic Cell Sorters is far from optimum for practical applications. Here we report a high-throughput Cell sorting method based on standing surface acoustic waves (SSAWs). We utilized a pair of focused interdigital transducers (FIDTs) to generate SSAW with high resolution and high energy efficiency. As a result, the sorting throughput is improved significantly from conventional acoustic-based Cell sorting methods. We demonstrated the successful sorting of 10 μm polystyrene particles with a minimum actuation time of 72 μs, which translates to a potential sorting rate of more than 13 800 events per second. Without using a Cell-detection unit, we were able to demonstrate an actual sorting throughput of 3300 events per second. Our sorting method can be conveniently integrated with upstream detection units, and it represents an important development towards a functional acoustic-based FACS system.

  • Acoustofluidic Fluorescence Activated Cell Sorter
    Analytical Chemistry, 2015
    Co-Authors: Ahmad Ahsan Nawaz, Yuchao Chen, Nitesh Nama, Ruth H. Nissly, Liqiang Ren, Adem Ozcelik, Lin Wang, J. Philip Mccoy, Stewart J. Levine, Tony Jun Huang
    Abstract:

    Selective isolation of Cell subpopulations with defined biological characteristics is crucial for many biological studies and clinical applications. In this work, we present the development of an acoustofluidic fluorescence activated Cell sorting (FACS) device that simultaneously performs on-demand, high-throughput, high-resolution Cell detection and sorting, integrated onto a single chip. Our acoustofluidic FACS device uses the "microfluidic drifting" technique to precisely focus Cells/particles three dimensionally and achieves a flow of single-file particles/Cells as they pass through a laser interrogation region. We then utilize short bursts (150 μs) of standing surface acoustic waves (SSAW) triggered by an electronic feedback system to sort fluorescently labeled particles/Cells with desired biological properties. We have demonstrated continuous isolation of fluorescently labeled HeLa Cells from unlabeled Cells at a throughput of ∼1200 events/s with a purity reaching 92.3 ± 3.39%. Furthermore, 99.18% postsort Cell viability indicates that our acoustofluidic sorting technique maintains a high integrity of Cells. Therefore, our integrated acoustofluidic FACS device is demonstrated to achieve two-way Cell sorting with high purity, biocompatibility, and biosafety. We believe that our device has significant potential for use as a low-cost, high-performance, portable, and user-friendly FACS instrument.

Yue Chen - One of the best experts on this subject based on the ideXlab platform.

  • pulsed laser activated Cell Sorter placs for high throughput fluorescent mammalian Cell sorting
    Proceedings of SPIE, 2014
    Co-Authors: Yue Chen, Michael A. Teitell, Dino Di Carlo, Aram J Chung, Yu Chung Kung, Pei-yu Chiou
    Abstract:

    We present a Pulsed Laser Activated Cell Sorter (PLACS) realized by exciting laser induced cavitation bubbles in a PDMS microfluidic channel to create high speed liquid jets to deflect detected fluorescent samples for high speed sorting. Pulse laser triggered cavitation bubbles can expand in few microseconds and provide a pressure higher than tens of MPa for fluid perturbation near the focused spot. This ultrafast switching mechanism has a complete on-off cycle less than 20 μsec. Two approaches have been utilized to achieve 3D sample focusing in PLACS. One is relying on multilayer PDMS channels to provide 3D hydrodynamic sheath flows. It offers accurate timing control of fast (2 m sec-1) passing particles so that synchronization with laser bubble excitation is possible, an critically important factor for high purity and high throughput sorting. PLACS with 3D hydrodynamic focusing is capable of sorting at 11,000 Cells/sec with >95% purity, and 45,000 Cells/sec with 45% purity using a single channel in a single step. We have also demonstrated 3D focusing using inertial flows in PLACS. This sheathless focusing approach requires 10 times lower initial Cell concentration than that in sheath-based focusing and avoids severe sample dilution from high volume sheath flows. Inertia PLACS is capable of sorting at 10,000 particles sec-1 with >90% sort purity.

  • 3d pulsed laser triggered high speed microfluidic fluorescence activated Cell Sorter
    Analyst, 2013
    Co-Authors: Yue Chen, Yu-chun Kung, Michael A. Teitell, Pei-yu Chiou
    Abstract:

    We report a 3D microfluidic pulsed laser-triggered fluorescence-activated Cell Sorter capable of sorting at a throughput of 23 000 Cells per s with 90% purity in high-purity mode and at a throughput of 45 000 Cells per s with 45% purity in enrichment mode in one stage and in a single channel. This performance is realized by exciting laser-induced cavitation bubbles in a 3D PDMS microfluidic channel to generate high-speed liquid jets that deflect detected fluorescent Cells and particles focused by 3D sheath flows. The ultrafast switching mechanism (20 μs complete on–off cycle), small liquid jet perturbation volume, and three-dimensional sheath flow focusing for accurate timing control of fast (1.5 m s−1) passing Cells and particles are three critical factors enabling high-purity sorting at high-throughput in this Sorter.

  • Pulsed laser triggered high speed microfluidic fluorescence activated Cell Sorter
    2012 IEEE 25th International Conference on Micro Electro Mechanical Systems (MEMS), 2012
    Co-Authors: Yue Chen, Michael A. Teitell, Dino Di Carlo, Sung Yong Park, Jason S. Hong, Tara Teslaa, Jiang F. Zhong, Pei-yu Chiou
    Abstract:

    We report a high speed and high purity pulsed laser triggered fluorescence activated Cell Sorter (PLACS) with a sorting throughput up to 20,000 mammalian Cells/s with 37% sorting purity, 90% Cell viability in enrichment mode, and >90% purity in high purity mode at 1,500 Cells/s or 3,000 beads/s. Fast switching (30 μs) and a small perturbation volume (∼90 pL) is realized by a unique sorting mechanism in which explosive vapor bubbles are generated using focused laser pulses in a single layer microfluidic PDMS channel.

  • 3d pulsed laser triggered high speed microfluidic fluorescence activated Cell Sorter
    Conference on Lasers and Electro-Optics, 2011
    Co-Authors: Yue Chen, Yu-chun Kung, Michael A. Teitell, Eric P Y Chiou
    Abstract:

    We report a high speed microfluidic fluorescence activated Cell Sorter (μFACS) capable of sorting at a throughput of 3000 beads/sec and 560 Cells/sec with >90% sample purity and 90% Cell viability after sorting. The sorting mechanism is realized by exciting dynamic vapor bubbles with focused laser pulses in a microfluidic PDMS channel. The explosive bubble expansion generates fast fluid flows which are directed into a neighboring sample channel to switch the particle of interest into the collection channel. This ultrafast laser triggered switching mechanism has the potential to advance the sorting speed of the state-of-the-art μFACS.