共查询到20条相似文献，搜索用时 0 毫秒
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《Small Methods》2018,2(1)
Considerable advances have been witnessed in the development of biochips that seek to realize various types of immune cell analysis on microscale platforms and enhance both basic and applied immunological research beyond the capability of conventional methods. Here, state‐of‐the‐art designs and examples for biochip‐based analysis and manipulation of immune cells are reviewed, and the potential of this emerging technology to enhance the understanding of immunology and improve disease diagnosis and treatment is discussed. In particular, some of the recent advances in this field, along with the challenges that must be addressed for these technologies, and their potential in precision medicine are highlighted. 相似文献
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Ken‐ichiro Kamei Yasumasa Mashimo Momoko Yoshioka Yumie Tokunaga Christopher Fockenberg Shiho Terada Yoshie Koyama Minako Nakajima Teiko Shibata‐Seki Li Liu Toshihiro Akaike Eiry Kobatake Siew‐Eng How Motonari Uesugi Yong Chen 《Small (Weinheim an der Bergstrasse, Germany)》2017,13(18)
Cellular microenvironments are generally sophisticated, but crucial for regulating the functions of human pluripotent stem cells (hPSCs). Despite tremendous effort in this field, the correlation between the environmental factors—especially the extracellular matrix and soluble cell factors—and the desired cellular functions remains largely unknown because of the lack of appropriate tools to recapitulate in vivo conditions and/or simultaneously evaluate the interplay of different environment factors. Here, a combinatorial platform is developed with integrated microfluidic channels and nanofibers, associated with a method of high‐content single‐cell analysis, to study the effects of environmental factors on stem cell phenotype. Particular attention is paid to the dependence of hPSC short‐term self‐renewal on the density and composition of extracellular matrices and initial cell seeding densities. Thus, this combinatorial approach provides insights into the underlying chemical and physical mechanisms that govern stem cell fate decisions. 相似文献
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Eduardo Reátegui Nicola Aceto Eugene J. Lim James P. Sullivan Anne E. Jensen Mahnaz Zeinali Joseph M. Martel Alexander J. Aranyosi Wei Li Steven Castleberry Aditya Bardia Lecia V. Sequist Daniel A. Haber Shyamala Maheswaran Paula T. Hammond Mehmet Toner Shannon L. Stott 《Advanced materials (Deerfield Beach, Fla.)》2015,27(9):1593-1599
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Mahlet Fasil Abate Shasha Jia Metages Gashaw Ahmed Xingrui Li Li Lin Xiaoqian Chen Zhi Zhu Chaoyong Yang 《Small (Weinheim an der Bergstrasse, Germany)》2019,15(14)
Immunocytological technologies, molecular technologies, and functional assays are widely used for detecting circulating tumor cells (CTCs) after enrichment from patients' blood sample. Unfortunately, accessibility to these technologies is limited due to the need for sophisticated instrumentation and skilled operators. Portable microfluidic devices have become attractive tools for expanding the access and efficiency of detection beyond hospitals to sites near the patient. Herein, a volumetric bar chart chip (V‐Chip) is developed as a portable platform for CTC detection. The target CTCs are labeled with aptamer‐conjugated nanoparticles (ACNPs) and analyzed by V‐Chip through quantifying the byproduct (oxygen) of the catalytic reaction between ACNPs and hydrogen peroxide, which results in the movement of an ink bar to a concentration‐dependent distance for visual quantitative readout. Thus, the CTC number is decoded into visually quantifiable information and a linear correlation can be found between the distance moved by the ink and number of cells in the sample. This method is sensitive enough that a single cell can be detected. Furthermore, the clinical capabilities of this system are demonstrated for quantitative CTC detection in the presence of a high leukocyte background. This portable detection method shows great potential for quantification of rare cells with single‐cell sensitivity for various applications. 相似文献
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Lang Nan Man Yuk Alison Lai Matthew Yuk Heng Tang Yau Kei Chan Leo Lit Man Poon Ho Cheung Shum 《Small (Weinheim an der Bergstrasse, Germany)》2020,16(9)
Droplet‐based microfluidic techniques are extensively used in efficient manipulation and genome‐wide analysis of individual cells, probing the heterogeneity among populations of individuals. However, the extraction and isolation of single cells from individual droplets remains difficult due to the inevitable sample loss during processing. Herein, an automated system for accurate collection of defined numbers of droplets containing single cells is presented. Based on alternate sorting and dispensing in three branch channels, the droplet number can be precisely controlled down to single‐droplet resolution. While encapsulating single cells and reserving one branch as a waste channel, sorting can be seamlessly integrated to enable on‐demand collection of single cells. Combined with a lossless recovery strategy, this technique achieves capture and culture of individual cells with a harvest rate of over 95%. The on‐demand droplet collection technique has great potential to realize quantitative processing and analysis of single cells for elucidating the role of cell‐to‐cell variations. 相似文献
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Kenza Samlali Fatemeh Ahmadi Angela B. V. Quach Guy Soffer Steve C. C. Shih 《Small (Weinheim an der Bergstrasse, Germany)》2020,16(34)
Generating a stable knockout cell line is a complex process that can take several months to complete. In this work, a microfluidic method that is capable of isolating single cells in droplets, selecting successful edited clones, and expansion of these isoclones is introduced. Using a hybrid microfluidics method, droplets in channels can be individually addressed using a co‐planar electrode system. In the hybrid microfluidics device, it is shown that single cells can be trapped and subsequently encapsulate them on demand into pL‐sized droplets. Furthermore, droplets containing single cells are either released, kept in the traps, or merged with other droplets by the application of an electric potential to the electrodes that is actuated through an in‐house user interface. This high precision control is used to successfully sort and recover single isoclones to establish monoclonal cell lines, which is demonstrated with a heterozygous NCI‐H1299 lung squamous cell population resulting from loss‐of‐function eGFP and RAF1 gene knockout transfections. 相似文献
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单细胞测序是指在单个细胞的水平上,对其相关组学进行测序分析的一项新技术。单细胞测序不仅能够分析单个细胞蕴含的遗传信息,还能综合评价多细胞异质性的问题,具有广阔的应用前景。在过去的十几年间,单细胞的研究从开始的单个细胞分离、扩增测序,发展到现今的高通量单细胞测序,陆续出现了众多新型、高效率的方法以及平台。综述了单细胞测序的发展进程,讨论了不同方法以及商用平台的优缺点及其适用性,并展望了单细胞测序对生物计量的影响。 相似文献
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Xuan Li Mohammad Aghaamoo Shiyue Liu Do‐Hyun Lee Abraham P. Lee 《Small (Weinheim an der Bergstrasse, Germany)》2018,14(40)
While lipoplex (cationic lipid‐nucleic acid complex)‐mediated intracellular delivery is widely adopted in mammalian cell transfection, its transfection efficiency for suspension cells, e.g., lymphatic and hematopoietic cells, is reported at only ≈5% or even lower. Here, efficient and consistent lipoplex‐mediated transfection is demonstrated for hard‐to‐transfect suspension cells via a single‐cell, droplet‐microfluidics approach. In these microdroplets, monodisperse lipoplexes for effective gene delivery are generated via chaotic mixing induced by the serpentine microchannel and co‐confined with single cells. Moreover, the cell membrane permeability increases due to the shear stress exerted on the single cells when they pass through the droplet pinch‐off junction. The transfection efficiency, examined by the delivery of the pcDNA3‐EGFP plasmid, improves from ≈5% to ≈50% for all three tested suspension cell lines, i.e., K562, THP‐1, Jurkat, and with significantly reduced cell‐to‐cell variation, compared to the bulk method. Efficient targeted knockout of the TP53BP1 gene for K562 cells via the CRISPR (clustered regularly interspaced short palindromic repeats)–CAS9 (CRISPR‐associated nuclease 9) mechanism is also achieved using this platform. Lipoplex‐mediated single‐cell transfection via droplet microfluidics is expected to have broad applications in gene therapy and regenerative medicine by providing high transfection efficiency and low cell‐to‐cell variation for hard‐to‐transfect suspension cells. 相似文献
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Xiaobao Cao Ying Du Andreas Küffner Jordan Van Wyk Paolo Arosio Jing Wang Peter Fischer Stavros Stavrakis Andrew deMello 《Small (Weinheim an der Bergstrasse, Germany)》2020,16(20)
Fluorescence‐based detection schemes provide for multiparameter analysis in a broad range of applications in the chemical and biological sciences. Toward the realization of fully portable analysis systems, microfluidic devices integrating diverse functional components have been implemented in a range of out‐of‐lab environments. That said, there still exits an unmet and recognized need for miniaturized, low‐cost, and sensitive optical detection systems, which provide not only for efficient molecular excitation, but also enhanced photon collection capabilities. To this end, an optofluidic platform that is adept at enhancing fluorescence light collection from microfluidic channels is presented. The central component of the detection module is a monolithic parabolic mirror located directly above the microfluidic channel, which acts to enhance the number of emitted photons reflected toward the detector. In addition, two‐photon polymerization is used to print a microscale‐lens below the microfluidic flow channel and directly opposite the mirror, to enhance the delivery of excitation radiation into the channel. Using such an approach, it is demonstrated that fluorescence signals can be enhanced by over two orders of magnitude, with component parallelization enabling the detection of pL‐volume droplets at rates up to 40 000 droplets per second. 相似文献
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Xuan Mu Wenfu Zheng Jiashu Sun Wei Zhang Xingyu Jiang 《Small (Weinheim an der Bergstrasse, Germany)》2013,9(1):9-21
Microfluidics, a toolbox comprising methods for precise manipulation of fluids at small length scales (micrometers to millimeters), has become useful for manipulating cells. Its uses range from dynamic management of cellular interactions to high‐throughput screening of cells, and to precise analysis of chemical contents in single cells. Microfluidics demonstrates a completely new perspective and an excellent practical way to manipulate cells for solving various needs in biology and medicine. This review introduces and comments on recent achievements and challenges of using microfluidics to manipulate and analyze cells. It is believed that microfluidics will assume an even greater role in the mechanistic understanding of cell biology and, eventually, in clinical applications. 相似文献
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Yizhi Zhang Zhuyuan Wang Lei Wu Shenfei Zong Binfeng Yun Yiping Cui 《Small (Weinheim an der Bergstrasse, Germany)》2018,14(20)
Isolating and in situ profiling the heterogeneous molecular phenotype of circulating tumor cells are of great significance for clinical cancer diagnosis and personalized therapy. Herein, an on‐chip strategy is proposed that combines size‐based microfluidic cell isolation with multiple spectrally orthogonal surface‐enhanced Raman spectroscopy (SERS) analysis for in situ profiling of cell membrane proteins and identification of cancer subpopulations. With the developed microfluidic chip, tumor cells are sieved from blood on the basis of size discrepancy. To enable multiplex phenotypic analysis, three kinds of spectrally orthogonal SERS aptamer nanovectors are designed, providing individual cells with composite spectral signatures in accordance with surface protein expression. Next, to statistically demultiplex the complex SERS signature and profile the cellular proteomic phenotype, a revised classic least square algorithm is employed to obtain the 3D phenotypic information at single‐cell resolution. Combined with categorization algorithm partial least square discriminate analysis, cells from different human breast cancer subtypes can be reliably classified with high sensitivity and selectivity. The results demonstrate that this platform can identify cancer subtypes with the spectral information correlated to the clinically relevant surface receptors, which holds great potential for clinical cancer diagnosis and precision medicine. 相似文献
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Tae Min Choi Gun Ho Lee Young‐Seok Kim Jin‐Gyu Park Hyerim Hwang Shin‐Hyun Kim 《Advanced materials (Deerfield Beach, Fla.)》2019,31(18)
Colloidal particles with a repulsive interparticle potential spontaneously form crystalline lattices, which are used as a motif for photonic materials. It is difficult to predict the crystal arrangement in spherical volume as lattices are incompatible with a spherical surface. Here, the optimum arrangement of charged colloids is experimentally investigated by encapsulating them in double‐emulsion drops. Under conditions of strong interparticle repulsion, the colloidal crystal rapidly grows from the surface toward the center of the microcapsule, forming an onion‐like arrangement. By contrast, for weak repulsion, crystallites slowly grow and fuse through rearrangement to form a single‐crystal phase. Single‐crystal structure is energetically favorable even for strong repulsion. Nevertheless, a high energy barrier to colloidal rearrangement kinetically arrests the onion‐like structure formed by heterogeneous nucleation. Unlike the isotropic onion‐shaped product, the anisotropic single‐crystal‐containing microcapsules selectively display—at certain orientations but not others—one of the distinct colors from the various crystal planes. 相似文献