一种可以实现稳定单细胞包裹的无进样器的微流控平台

doi:10.7523/j.issn.2095-6134.2020.03.006

中国科学院大学学报 ›› 2020, Vol. 37 ›› Issue (3): 336-344.DOI: 10.7523/j.issn.2095-6134.2020.03.006

一种可以实现稳定单细胞包裹的无进样器的微流控平台

范蓓媛^1,2, 刘力行^1,2, 李秀锋^1,2, 陈德勇^1,2, 王文会³, 王军波^1,2, 陈健^1,2

1. 中国科学院电子学研究所传感技术国家重点实验室, 北京 100190;
2. 中国科学院大学, 北京 100049;
3. 清华大学精密仪器系, 北京 100084

收稿日期:2018-10-10 修回日期:2019-03-04 发布日期:2020-05-15
通讯作者: 王军波, 陈健
基金资助:
Supported by the National Basic Research Program of China (2014CB744600), National Natural Science Foundation of China (61431019, 61671430), Chinese Academy of Sciences Key Project Targeting Cutting-Edge Scientific Problems (QYZDB-SSW-JSC011), Instrument Development Program of the Chinese Academy of Sciences and Youth Innovation Promotion Association of Chinese Academy of Sciences

Study of syringe-free droplet microfluidic platforms enabling stable single-cell encapsulation

FAN Beiyuan^1,2, LIU Lixing^1,2, LI Xiufeng^1,2, CHEN Deyong^1,2, WANG Wenhui³, WANG Junbo^1,2, CHEN Jian^1,2

1. . State Key Laboratory of Transducer Technology, Institute of Electronics, Chinese Academy of Sciences, Beijing 100190, China;
2. . University of Chinese Academy of Sciences, Beijing 100049, China;
3. . Department of Precision Instrument, Tsinghua University, Beijing 100084, China

Received:2018-10-10 Revised:2019-03-04 Published:2020-05-15
Supported by:
Supported by the National Basic Research Program of China (2014CB744600), National Natural Science Foundation of China (61431019, 61671430), Chinese Academy of Sciences Key Project Targeting Cutting-Edge Scientific Problems (QYZDB-SSW-JSC011), Instrument Development Program of the Chinese Academy of Sciences and Youth Innovation Promotion Association of Chinese Academy of Sciences

摘要/Abstract

摘要： 液滴单细胞包裹技术是实现单细胞研究的有效方法。采用负压作为驱动力包裹单细胞，不再需要常规实验中会引起细胞吸附和降低单细胞包裹率的进样器和管路。在细胞悬浮溶液中加入碘克沙醇溶液调整溶液密度，进一步改善细胞沉降问题。研究负压大小和T型沟道几何尺寸对液滴频率和体积的影响以及碘克沙醇调整溶液密度后对单细胞包裹率的影响。该微流控平台可实现稳定的单细胞包裹，并为单细胞分析提供有利工具。

关键词: 液滴微流控, 单细胞包裹, 负压, 碘克沙醇

Abstract: Negative pressures were used to generate droplets for single-cell encapsulations. Iodixanol was used to change the suspension solution densities in order to reduce the issue of cellular sedimentations. The effects of negative pressures and geometries of microfluidic T-channels on volumes and frequencies of generated droplets and the effects of iodixanol on the percentages of single-cell encapsulations were investigated and compared. This microfluidic platform may enable stable single-cell encapsulations, paving ways for single-cell analysis.

Key words: droplet microfluidics, single-cell encapsulation, negative pressure, iodixanol

中图分类号:

范蓓媛, 刘力行, 李秀锋, 陈德勇, 王文会, 王军波, 陈健. 一种可以实现稳定单细胞包裹的无进样器的微流控平台[J]. 中国科学院大学学报, 2020, 37(3): 336-344.

FAN Beiyuan, LIU Lixing, LI Xiufeng, CHEN Deyong, WANG Wenhui, WANG Junbo, CHEN Jian. Study of syringe-free droplet microfluidic platforms enabling stable single-cell encapsulation[J]. , 2020, 37(3): 336-344.

参考文献

[1] Christopher G F, Anna S L. Microfluidic methods for generating continuous droplet streams[J]. Journal of Physics D:Applied Physics, 2007, 40(19):R319-R336.
[2] Teh S Y, Lin R, Hung L H, et al. Droplet microfluidics[J]. Lab Chip, 2008, 8(2):198-220.
[3] Ralf S, Martin B, Thomas P, et al. Droplet based microfluidics[J]. Reports on Progress in Physics, 2012, 75(1):016601.
[4] Leman M, Abouakil F, Griffiths A D, et al. Droplet-based microfluidics at the femtolitre scale[J]. Lab on a Chip, 2015, 15(3):53-65.
[5] Theberge A B, Courtois F, Schaerli Y, et al. Microdroplets in microfluidics:an evolving platform for discoveries in chemistry and biology[J]. Angewandte Chemie International Edition, 2010, 49(34):5846-5868.
[6] Basova E Y, Foret F. Droplet microfluidics in (bio)chemical analysis[J]. Analyst, 2015, 140(1):22-38.
[7] Shembekar N, Chaipan C, Utharala R, et al. Droplet-based microfluidics in drug discovery, transcriptomics and high-throughput molecular genetics[J]. Lab on a Chip, 2016, 16:1314-1331.
[8] Brouzes E. Droplet microfluidics for single-cell analysis[J]. Methods in Molecular Biology, 2012, 853:105-139.
[9] Joensson H, Andersson Svahn H. Droplet microfluidics-a tool for single-cell analysis[J]. Angewandte Chemie International Edition, 2012, 51(49):12176-12192.
[10] Collins D J, Neild A, DeMello A, et al. The Poisson distribution and beyond:methods for microfluidic droplet production and single cell encapsulation[J]. Lab Chip, 2015, 15(17):3439-3459.
[11] Wen N, Zhao Z, Fan B, et al. Development of droplet microfluidics enabling high-throughput single-cell analysis[J]. Molecules, 2016, 21(7):881.
[12] He M, Edgar J S, Jeffries G D, et al. Selective encapsulation of single cells and subcellular organelles into picoliter- and femtoliter-volume droplets[J]. Analytical Chemistry, 2005, 77(6):1539-1544.
[13] Tan Y C, Hettiarachchi K, Siu M, et al. Controlled microfluidic encapsulation of cells, proteins, and microbeads in lipid vesicles[J]. Journal of the American Chemical Society, 2006, 128(17):5656-5658.
[14] Luo D, Pullela S R, Marquez M, et al. Cell encapsules with tunable transport and mechanical properties[J]. Biomicrofluidics, 2007, 1(3):34102.
[15] Zhu L, Peh X L, Ji H M, et al. Cell loss in integrated microfluidic device[J]. Biomed Microdevices, Oct 2007, 9(5):745-750.
[16] Chabert M, Viovy J M. Microfluidic high-throughput encapsulation and hydrodynamic self-sorting of single cells[J]. Proceedings of the National Academy of Sciences of the United States of America, Mar 42008, 105(9):3191-3196.
[17] Baret J C, Beck Y, Billas-Massobrio I, et al. Quantitative cell-based reporter gene assays using droplet-based microfluidics[J]. Chemistry and Biology, 2010, 17(5):528-536.
[18] Clausell-Tormos J, Lieber D, Baret J C, et al. Droplet-based microfluidic platforms for the encapsulation and screening of Mammalian cells and multicellular organisms[J]. Chemistry & Biology, 2008, 15(5):427-437.
[19] Kemna E W M, Schoeman R M, Wolbers F, et al. High-yield cell ordering and deterministic cell-in-droplet encapsulation using Dean flow in a curved microchannel[J]. Lab Chip, 2012, 12:2881-2887.
[20] Lagus T P, Edd J F. High throughput single-cell and multiple-cell micro-encapsulation[J]. Journal of Visualized Experiments:JoVE, 2012, 64:e4096.
[21] Mazutis L, Gilbert J, Ung W L, et al. Single-cell analysis and sorting using droplet-based microfluidics[J]. Nature Protocols, 2013, 8:870-891.
[22] Kintses B, Hein C, Mohamed M F, et al. Picoliter cell lysate assays in microfluidic droplet compartments for directed enzyme evolution[J]. Chemistry and Biology, 2012, 19(8):1001-1009.
[23] Koster S, Angile F E, Duan H, et al. Drop-based microfluidic devices for encapsulation of single cells[J]. Lab Chip, 2008, 8(7):1110-1115.
[24] Frenz L, Blank K, Brouzes E, et al. Griffiths. Reliable microfluidic on-chip incubation of droplets in delay-lines[J]. Lab Chip, 2009, 9:1344-1348.
[25] Huebner A, Srisa-Art M, Hol D, et al. Quantitative detection of protein expression in single cells using droplet microfluidics[J]. Chemical Communications, 2007, 12:1218-1220.
[26] Fidalgo L M, Abell C, Huck W T. Surface-induced droplet fusion in microfluidic devices[J]. Lab Chip, 2007, 7(8):984-986.
[27] Baret J C, Miller O J, Taly V, et al. Fluorescence-activated droplet sorting (FADS):efficient microfluidic cell sorting based on enzymatic activity[J]. Lab on a Chip, 2009, 9:1850-1858.
[28] Shintaku H, Kuwabara T, Kawano S, et al. Micro cell encapsulation and its hydrogel-beads production using microfluidic device[J]. Microsystem Technologies, 2007, 13(8-10):951-958.
[29] Li H, Xue Y, Xu M, et al. Viscosity based droplet size controlling in negative pressure driven droplets generator for large-scale particle synthesis[J]. Electrophoresis, 2017, 38(13/14):1736-1742.
[30] Teo A J T, Li K H, Nguyen N T, et al. Negative pressure induced droplet generation in a microfluidic flow-focusing device[J]. Analytical Chemistry, 2017, 89(8):4387-4391.

一种可以实现稳定单细胞包裹的无进样器的微流控平台

Study of syringe-free droplet microfluidic platforms enabling stable single-cell encapsulation

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