Microfluidics for size and deformability based cell sorting

MICROFLUIDICS FOR SIZE AND DEFORMABILITY BASED CELL SORTING GUAN GUOFENG A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF MECHANICAL ENGINEERING NATIONAL UNIVERSITY OF SINGAPORE 2013 DECLARATION I hereby declare that this thesis is my orginal work and it has been written by me in its entirely. I have duly acknowledged all the sources of information which have been used in this thesis. This thesis has also not been submitted for any degree in any university previously. Guofeng Guan 20 January 2013 i Acknowledgements This thesis would not have been possible without the guidance and support of many people who in one way or another contributed and extended their valuable assistance in the preparation and completion of this study. I am heartily thankful to my supervisor, Dr. Peter Chen Chao Yu, my cosupervisor, Dr. Chong Jin Ong from Department of Mechanical Engineering, National University of Singapore, and my research PI in Singapore MIT Alliance for Research and Technology (SMART) Centre's BioSyM IRG, Dr. Jongyoon Han from Department of Biological Engineering, Massachusetts Institute of Technology for their invaluable encouragement, enthusiasm and guidance from the initial to the final level of this project. This thesis would not have been successful without their knowledge and support. I would like to express my appreciation to Dr. Ali Asgar Bhagat, Dr. Brain Weng Kung Peng, Dr. Majid Ebrahimi Warkiani, Prof. Zirui Li, and all other Post-doctoral and graduate students from SMART BioSyM IRG, for sharing their knowledge and invaluable assistance. Special thanks also to Dr. Narayanan Balasubramanian, Mr. chee Keong Kwok and all others staffs from Singapore MIT Alliance for Research and Technology, for their kindly assistance. Many thanks to the examiners, especially Dr. Teo Chiang Juay, who’s careful review and comments greatly improved the quality of the thesis. Last but not the least, I wish to thank all my fellow colleagues, especially group members, Mr. Shengfeng Zhou, Mr. Sahan Christie Bandara Herath, Miss. Yue Du and all the staffs from Control and Mechatronics Lab, for their friendship, assistance and kindness. Finally, I would like to acknowledge the National University of Singapore for the financial support in the form of a Research Scholarship and the financial support of National Research Foundation Singapore, through SMART BioSyM IRG research programme for the study. ii Contents List of Tables viii List of Figures ix List of Symbols and Acronyms xvi Introduction 1.1 Biophysical and Biomechanical Properties as Label-free Cell Markers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.1.1 Mesenchymal Stem Cells . . . . . . . . . . . . . . . . . . 1.1.2 Cancer and Circulating Tumor Cells . . . . . . . . . . . . 1.1.3 Malaria . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.2 The Requiremnts of New Technology for Lab-On-a-Chip, Size and Deformability Based Cell Sorting . . . . . . . . . . . . . . . 1.3 Objective of the New Size and Deformability Based Cell Sorting Methods Development . . . . . . . . . . . . . . . . . . . . . . . 1.4 Organization of the Thesis . . . . . . . . . . . . . . . . . . . . . Background and Literature Review 2.1 Microfluidics Methodologies for Size Based Cell Separation . 2.1.1 Active Separation Methods with External Force Fields 2.1.2 Passive Separation Methods . . . . . . . . . . . . . . 2.2 Methodologies for Deformability Based Cell Sorting . . . . . 2.3 Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Mechanism of Dean-Inertial Microfluidics for Particle Focusing 3.1 Experimental Observation of Side View Particle Focusing in Spiral Microfluidic Channel . . . . . . . . . . . . . . . . . . . . . . 3.1.1 Device design and fabrication . . . . . . . . . . . . . . . 3.1.2 Fluid preparation . . . . . . . . . . . . . . . . . . . . . . 3.1.3 Result of Side View Focusing . . . . . . . . . . . . . . . iii 2 8 26 34 36 37 37 38 39 CONTENTS 3.2 3.3 3.4 Simulation and Force Analysis . . . . . . . . . . . . . . . . . . . 3.2.1 Numerical Simulation of Dean Flow Field . . . . . . . . . 3.2.2 Force balance analysis of particle in curved channel . . . . Particle Focusing and Migration Process in Curved Rectangular cross section Microfluidic Channel . . . . . . . . . . . . . . . . . Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . Trapezoidal cross section spiral microfluidics for size based particle separation 4.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.2 Particle Focusing Positions in Trapezoidal Cross Section Spiral Channel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.2.1 3D Observation . . . . . . . . . . . . . . . . . . . . . . . 4.2.2 Comparison of top view focusing . . . . . . . . . . . . . 4.2.3 Force Analysis . . . . . . . . . . . . . . . . . . . . . . . 4.3 Separation Resolution and Throughput . . . . . . . . . . . . . . . 4.3.1 The effect of geometry of channel cross section . . . . . . 4.4 Sorting of Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . 4.4.1 Cyclic Cell Sorting . . . . . . . . . . . . . . . . . . . . . 4.4.2 Blood Cells . . . . . . . . . . . . . . . . . . . . . . . . . 4.4.3 Mesenchymal Stem Cells . . . . . . . . . . . . . . . . . . 4.5 Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . 41 41 43 50 53 55 55 57 57 58 61 62 65 68 69 69 71 74 Size-Independent Deformability Sorting with Real-time Controlled Microfluidic Channel 76 5.1 Introduction of the Approach . . . . . . . . . . . . . . . . . . . . 77 5.2 Model of the Cell and Squeezing Process . . . . . . . . . . . . . 79 5.2.1 Flow and Pressure Impedance inside Channel . . . . . . . 80 5.2.2 Model of the Cell and Squeezing Process . . . . . . . . . 83 5.3 Implementation of the System . . . . . . . . . . . . . . . . . . . 84 5.3.1 Design and fabrication of the microfluidic device with a controllable channel . . . . . . . . . . . . . . . . . . . . 84 5.3.2 Calibration and characterization of the control channel . . 86 5.3.3 Cell imaging, data collection and processing . . . . . . . 87 5.3.4 Real-time control of channel gap . . . . . . . . . . . . . . 89 5.4 Experiments and Results for Cell Sorting . . . . . . . . . . . . . 90 5.4.1 MCF-7 and MCF-10A . . . . . . . . . . . . . . . . . . . 90 5.4.2 Mesenchymal stem cells . . . . . . . . . . . . . . . . . . 93 5.4.3 Error analysis . . . . . . . . . . . . . . . . . . . . . . . . 94 5.5 Concluding Remarks . . . . . . . . . . . . . . . . . . . . . . . . 95 iv CONTENTS Conclusion and Future Direction 97 References 104 v Summary Label free cell sorting and separation is a new field in biomedical and chemical analysis. Many devices and technologies have been developed in recent years utilizing cell properties such as size and deformability. Although attempts have been made to increase the throughput and efficiency of sorting, there is still a significant gap from lab experiments to clinical application. The aim of this study was to investigate cell sorting and separation technologies with respect to both cell size and deformability experimently and theoretically. Using polydimethylsiloxane (PDMS) microfluidic technologies, new devices for size- and deformability-based cell sorting were designed and their performance studied. Firstly, for developing high throughput size-based separation method, threedimensional observation of the location of focused particle streams along both the height and width of the channel cross-section in spiral inertial microfluidic systems was proposed. The results confirmed that particles are focused near the top and bottom wall of microchannel cross-section, revealing clear insights regarding the balance of forces acting on the particles. Based on this detailed understanding of the force balance, a novel spiral microchannel with trapezoidal cross section that generate stronger Dean vortices at the outer half of the channel was developed. Experiments show that the focusing position of particles in such a device is sensitive to particle size and flow rate, and exhibits a sharp transition from the inner half to the outer half of the equilibrium positions at a size-dependent critical flow rate. As particle equilibration positions are well segregated based on different focusing mechanisms, higher separation resolution was achieved in such a system over conventional spiral microchannels with rectangular cross-section. Further studies with particles and cells indicated that this channel is able to handle sample with particle concentration of up to 1.8%. The separation results on Mesenchymal stem cells (MSCs) indicated that cell deformability might influence the separation efficiency. In this case, the deformability should be considered as a secondary bio-marker. vi SUMMARY A new microfluidic system with real-time feedback control to evaluate single cell deformability while minimizing cell-size dependence of the measurement was thus developed. The system consists of a microfluidic chip with two cross channels, i.e., the control channel and the flow channel, forming a membrane area in between. By adjusting the pressure in the control channel, the deformed membrane can generate a controllable bottleneck section in the flow channel. The bottleneck was used for measuring the deformability of cells by adjusting the height of the bottleneck in real time according to the diameter of the cells. The influence of size variation can thus be eliminated during the measurement. Using breast cancer cells (MCF-7), the potential of this system for stiffness-profiling of cells in a complex, diverse cell populations was demonstrated. The comparison of time spent for MCF-7 cells and MCF10A cells, a healthy breast cell line, to squeeze through the bottleneck section of the flow channel indicated that MCF-10A cells are much stiffer. This result confirmed reports from studies by other researchers. Mathematical models for both size- and deformability-based sorting were developed. For device with a spiral microfluidic channel, forces applied to particles inside both rectangular and trapezoidal channel cross-sections at various positions were analysed and discussed. Theses analysis provided a detailed explanation on the particle focusing mechanism in a curved microfluidic channel. For deformability-based sorting device, a mathematical model on a cell squeezing through a bottleneck channel section was constructed. Based on this model, the elastic and viscous properties of both MCF-7 and MCF-10A were quantified. vii List of Tables 2.1 2.2 Size based passive cell/particle sorting technologies . . . . . . . . Deformability based cell sorting technologies . . . . . . . . . . . viii 25 33 List of Figures 2.1 2.2 2.3 2.4 2.5 The schematic illustration of active separation with external force field. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Schematic illustrating size-exclusion separation designs. (A) Weir structure size-exclude cellular allowing flow of smaller cells to pass through a planar slit. (B) Arrays of pillars which exclude cells larger than the spacing of the pillars. (C) Crescent-shaped isolation wells that only trap larger cells. (D) Cross-flow filters that alow continuous flow and collection of both large and small cells from different outlets. . . . . . . . . . . . . . . . . . . . . . Schematic illustrating the separation by deterministic lateral displacement in an array of microposts.The asymmetrical placements of the posts in the array cause particles of different sizes to follow different flow paths. This results in an in lateral displacement and thus separation of particles by size. Image reprinted with permission from [71], copyright 2008, by The National Academy of Sciences of the USA. . . . . . . . . . . . . . . . . . . . . . . . . Schematic illustrating the wall-against alignment based separation technologies. Cells/particles are aligned to the channel side wall(s) by (A) pushing the sample stream along side wall with a sheath buffer, or (B) draining liquid from both channel side branches. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . (A) Schematic of straight channel design with one inlet and three outlets for inertial focusing based blood separation. Figure reprinted with permission from [64], copyright 2010, Wiley Periodicals, Inc. (B) The concept of soft inertial separation with the formation of the curved and focused sample flow segment and particle momentum loss induced inertial force on fluid element. Figure reprinted with permission from [109], copyright 2009, The Royal Society of Chemistry. . . . . . . . . . . . . . . . . . . . . . . . . ix 10 11 13 14 15 Conclusion and Future Direction utilized. By controlling the deformation of the membrane via adjusting the pressure on the other side i.e. in the control channel with a pressure regulator, the channel height of this bottleneck section is dynamically set to be a fraction of the diameter of individual cells, the cells are forced to deform as they pass through the bottleneck section under hydraulic drag and channel wall press. The passage time can be online measured with optical microscope and used as an index for differentiating cells of various sizes by their deformability. The advantage this approach over the conventional method of using a fixedgap channel have been demonstrated using MCF-7 cell line. The effectiveness of this approach in differentiating cell populations of cells with inherent size variation by their deformability was further demonstrated by the comparison between MCF-7 and MCF-10A cell lines. The results are in line with former reports that MCF-10A cells are stiffer than MCF-7 cells by analysing the magnitude of the Young’s modulus [21], simply by the cell passage time [40], or other information as elongation [31, 60] or impedance [48]. By simplifying the problem of a cell squeezing through a bottleneck section as the deformation of a viscoelastic sphere under a constant pressing force, a mathematical model was built and the elastic modulus of both cell lines was given. There are several factors that affect the throughput of our deformability based sorting device. Since the release of cells from the inlet into the channel, and the movement of cells in the flow channel are not controlled on a per-cell basis, the throughput of the current system can be highly erratic, thus the concentration of cells flowing through the channel is limited. Moreover, the non-uniform distribution of cells in the flow may result in multiple cells entering the bottleneck section during one measurement cycle, leading to inaccuracy in the measurement data. One way to resolve this problem is to control the cell release at the inlet so as to maintain a per-cell flow rate that can accommodate the possible variation in the passage time of individual cells, such that only one cell is allowed to be in the bottleneck section during any given measurement cycle. The performance of the equipments also affect the sorting throughput in several different aspects. The time cost in online image processing and the response of pressure regulator are the key limitations. We believe that, with the development of the computing hardware and the pneumatic components, the throughput of the deformability based sorting 100 Conclusion and Future Direction device can be further increased. 101 List of Publications Journal Papers 1. Guofeng Guan, Lidan Wu, Ali Asgar Bhagat, Zirui Li, Peter C Y Chen, Shuzhe Chao, Chong Jin Ong, and Jongyoon Han, Spiral Microchannel with Rectangular and Trapezoidal Cross-sections for Size Based Particle Separation, Scientific Reports 3, (2013), 1475 2. Guofeng Guan, Peter C Y Chen, Weng Kung Peng, Ali Asgar Bhagat, Chong Jin Ong, and Jongyoon Han, Real-time Control of a Microfluidic Channel for Size-independent Deformability Cytometry, Journal of Micromechanics and Microengineering, 22 (2012), 105037 3. Lidan Wu, Guofeng Guan, Han Wei Hou, Ali Asgar S Bhagat, and Jongyoon Han, Separation of Leukocytes from Blood Using Spiral Channel with Trapezoid Cross-section., Analytical chemistry, 84 (2012), 9324-31 4. Rhokyun Kwak, Guofeng Guan, Weng Kung Peng, and Jongyoon Han, Microscale Electrodialysis: Concentration Profiling and Vortex Visualization, Desalination, 308 (2013), 138-46 5. Shuzhe Chao, Geok-Soon Hong, and Guofeng Guan, The Design and Field Test of a Micro AUV Lancelet with a Multi-jet Drive Propulsion System, Ocean Engineering (submitted) Conference Proceedings 1. Guofeng Guan, Ali Asgar Bhagat, Weng Kung Peng, Wong Cheng Lee, Chong Jin Ong, Peter C Y Chen, and Jongyoon Han, Size-Independent Deformability Cytometry with Active Feedback Control of Microfluidic Channels, in MicroTAS, 2011, 1053-5 2. Guofeng Guan, Ali Asgar Bhagat, Lidan Wu, Zirui Li, Chong Jin Ong, Peter C Y Chen, and Jongyoon Han, High Resolution Size Based Micro Particle/Cell Separator With Trapezoidal Cross Section Spiral Microchannels, in MicroTAS, 2012, 518-20 102 LIST OF PUBLICATIONS 3. Lidan Wu, Guofeng Guan, Han Wei Hou, Ali Asgar S Bhagat, and Jongyoon Han, Continuous RBS Removal Using Spiral Microchannel With Trapeziod Cross-Section, in MicroTAS, 2012, 1099-101 103 References [1] Emad A-Hassan, William F Heinz, Matthew D Antonik, Neill P D’Costa, Soni Nageswaran, Cora-Ann Schoenenberger, and Jan H Hoh. Relative microelastic mapping of living cells by atomic force microscopy. Biophysical Journal, 74(3):1564–78, March 1998. [2] Hamed Amini and Elodie Sollier. Intrinsic particle-induced lateral transport in microchannels. Proceedings of the National Academy of Sciences, 109(29):11593–8, 2012. [3] A Ashkin, J M Dziedzic, J E Bjorkholm, and Steven Chu. Observation of a single-beam gradient-force optical trap for dielectric particles in air. Optics letters, 11(5):288–90, June 1986. [4] Evgeny S. Asmolov. The inertial lift on a spherical particle in a plane Poiseuille flow at large channel Reynolds number. Journal of Fluid Mechanics, 381:63–87, 1999. [5] Mark Bathe, Atsushi Shirai, Claire M Doerschuk, and Roger D Kamm. Neutrophil transit times through pulmonary capillaries: the effects of capillary geometry and fMLP-stimulation. Biophysical journal, 83(4):1917–33, October 2002. [6] Jenny M Bator, Michael R Groves, Brandon J Price, and Eugene C Eckstein. Erythrocyte Deformability and Size Measured in a Multiparameter System That Includes Impedance Sizing. Cytometry, 41:34–41, 1984. [7] Andreas R Bausch, Winfried M¨oller, and Erich Sackmann. Measurement of local viscoelasticity and forces in living cells by magnetic tweezers. Biophysical Journal, 76:573–9, January 1999. [8] Ali Asgar S Bhagat, Han Wei Hou, Leon D Li, Chwee Teck Lim, and Jongyoon Han. Dean Flow Fractionation(DFF) Isolation of Circulating Tumor Cells (CTCs) From Blood. In microTAS, pages 524–526, 2011. 104 REFERENCES [9] Ali Asgar S Bhagat, Han Wei Hou, Leon D Li, Chwee Teck Lim, and Jongyoon Han. Pinched flow coupled shear-modulated inertial microfluidics for high-throughput rare blood cell separation. Lab on a chip, 11(11):1870–8, June 2011. [10] Ali Asgar S Bhagat, Sathyakumar S Kuntaegowdanahalli, Necati Kaval, Carl J Seliskar, and Ian Papautsky. Inertial microfluidics for sheath-less high-throughput flow cytometry. Biomedical microdevices, 12(2):187–95, April 2010. [11] Ali Asgar S Bhagat, Sathyakumar S Kuntaegowdanahalli, and Ian Papautsky. Continuous particle separation in spiral microchannels using Dean flows and differential migration. Lab on a chip, 8(11):1906–14, November 2008. [12] Ali Asgar S Bhagat, Sathyakumar S Kuntaegowdanahalli, and Ian Papautsky. Inertial microfluidics for continuous particle filtration and extraction. Microfluidics and Nanofluidics, 7(2):217–226, December 2008. [13] Hansen Bow, Igor V Pivkin, Monica Diez-silva, Stephen J Goldfless, Ming Dao, and Jacquin C Niles. A microfabricated deformability-based flow cytometer with application to malaria. Lab on a Chip, 11:1065–1073, 2011. [14] Hansen Chang Bow. Microfluidic Devices for Analysis of Red Blood Cell Mechanical Properties. PhD thesis, Massachusetts Institute of Technology, 2010. [15] James P Brody, Yuqi Han, Robert H Austin, and Mark Bitensky. Deformation and flow of red blood cells in a synthetic lattice: evidence for an active cytoskeleton. Biophysical Journal, 68(6):2224–2232, June 1995. [16] Andres M Cardenas-Valencia, Jay Dlutowski, David Fries, and Larry Langebrake. Spectrometric determination of the refractive index of optical wave guiding materials used in lab-on-a-chip applications. Applied spectroscopy, 60(3):322–9, March 2006. [17] Pradeep Cherukat and John B. McLaughlin. The inertial lift on a rigid sphere in a linear shear flow field near a flat wall. Journal of Fluid Mechanics, 263:1–18, 1994. [18] Choeng Ryul Choi and Chang Nyung Kim. Inertial migration and multiple equilibrium positions of a neutrally buoyant spherical particle in Poiseuille flow. Korean Journal of Chemical Engineering, 27(4):1076–1086, May 2010. 105 REFERENCES [19] Yong-seok Choi, Kyung-won Seo, and Sang-joon Lee. Lateral and crosslateral focusing of spherical particles in a square microchannel. Lab on a chip, 11(3):460–5, February 2011. [20] C F M Coimbra and R H Rangel. General solution of the particle momentum equation in unsteady Stokes flows. Journal of Fluid Mechanics, 370(July 2002):53–72, September 1998. [21] Sarah E Cross, Yu-Sheng Jin, Jianyu Rao, and James K Gimzewski. Nanomechanical analysis of cells from cancer patients. Nature nanotechnology, 2(12):780–3, December 2007. [22] Sarah E Cross, Yu-Sheng Jin, Julianne Tondre, Roger Wong, Jian Yu Rao, and James K Gimzewski. AFM-based analysis of human metastatic cancer cells. Nanotechnology, 19(38):384003, September 2008. [23] Dino Di Carlo. Inertial microfluidics. Lab on a chip, 9(21):3038–46, November 2009. [24] Dino Di Carlo, Jon Edd, Katherine Humphry, Howard Stone, and Mehmet Toner. Particle Segregation and Dynamics in Confined Flows. Physical Review Letters, 102(9):1–4, March 2009. [25] Dino Di Carlo, Jon F Edd, Daniel Irimia, Ronald G Tompkins, and Mehmet Toner. Equilibrium separation and filtration of particles using differential inertial focusing. Analytical chemistry, 80(6):2204–11, March 2008. [26] Dino Di Carlo, Daniel Irimia, Ronald G Tompkins, and Mehmet Toner. Continuous inertial focusing, ordering, and separation of particles in microchannels. Proceedings of the National Academy of Sciences of the United States of America, 104(48):18892–7, November 2007. [27] A El Hasni, K G¨obbels, A L Thiebes, P Br¨aunig, W Mokwa, and U Schnakenberg. Focusing and Sorting of Particles in Spiral Microfluidic Channels. Procedia Engineering, 25(0):1197–1200, January 2011. [28] Shady Gawad, Karen Cheung, Urban Seger, Arnaud Bertsch, and Philippe Renaud. Dielectric spectroscopy in a micromachined flow cytometer: theoretical and practical considerations. Lab on a chip, 4(3):241–51, June 2004. [29] Sean C Gifford, Michael G Frank, Jure Derganc, Christopher Gabel, Robert H Austin, Tatsuro Yoshida, and Mark W Bitensky. Parallel microchannel-based measurements of individual erythrocyte areas and volumes. Biophysical journal, 84(1):623–33, January 2003. 106 REFERENCES [30] Daniel R Gossett and Dino Di Carlo. Particle focusing mechanisms in curving confined flows. Analytical chemistry, 81(20):8459–65, October 2009. [31] Daniel R Gossett, Henry T K Tse, Serena A Lee, Amander T Clark, and D Di Carlo. Deformability cytometry: high-throughput,domtinuous measurement of cell mechanical properties in extesional flow. In microTAS, number October, pages 1382–1384, 2010. [32] Daniel R Gossett, Henry T K Tse, Serena A Lee, Yong Ying, Anne G Lindgren, Otto O Yang, Jianyu Rao, Amander T Clark, and Dino Di Carlo. Hydrodynamic stretching of single cells for large population mechanical phenotyping. Proceedings of the National Academy of Sciences of the United States of America, 109(20):7630–5, May 2012. [33] James V Green, Milica Radisic, and Shashi K Murthy. Deterministic lateral displacement as a means to enrich large cells for tissue engineering. Analytical chemistry, 81(21):9178–82, November 2009. [34] Jochen Guck, Stefan Schinkinger, Bryan Lincoln, Falk Wottawah, Susanne Ebert, Maren Romeyke, Dominik Lenz, Harold M Erickson, Revathi Ananthakrishnan, Daniel Mitchell, Josef K¨as, Sydney Ulvick, and Curt Bilby. Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. Biophysical journal, 88(5):3689–98, May 2005. [35] The Sydney Morning Herald. Gorillas in midst of malaria mystery. http: //www.who.int/mediacentre/factsheets/fs297/en/, February 2009. [36] Robert M Hochmuth. Micropipette aspiration of living cells. Journal of Biomechanics, 33(1):15–22, January 2000. [37] Danial N Hohne, John G Younger, and Michael J Solomon. Flexible microfluidic device for mechanical property characterization of soft viscoelastic solids such as bacterial biofilms. Langmuir, 25(13):7743–51, July 2009. [38] Stefan H Holm, Jason P Beech, Michael P Barrett, and Jonas O Tegenfeldt. Separation of parasites from human blood using deterministic lateral displacement. Lab on a chip, 11(7):1326–32, April 2011. [39] L H Bannisterand J M Hopkins, R E Fowler, Krishna S, and G H Mitchell. A brief illustrated guide to the ultrastructure of plasmodium falciparum asexual blood stages. Parasitol Today, 16:427433, 2000. 107 REFERENCES [40] Han Wei Hou, Q S Li, G Y H Lee, A P Kumar, C N Ong, and Chwee Teck Lim. Deformability study of breast cancer cells using microfluidics. Biomedical Microdevices, 11(3):557–64, June 2009. [41] Lotien Richard Huang, Edward C Cox, Robert H Austin, and James C Sturm. Continuous particle separation through deterministic lateral displacement. Science, 304(5673):987–90, May 2004. [42] R Huang, T A Barber, M A Schmidt, R G Tompkins, M Toner, D W Bianchi, R Kapur, and W L Flejter. A microfluidics approach for the isolation of nucleated red blood cells ( NRBCs ) from the peripheral blood of pregnant women. Prenatal Diagnosis, (May):892–899, 2008. [43] Soojung Claire Hur, Tatiana Z Brinckerhoff, Christopher M Walthers, James C Y Dunn, and Dino Di Carlo. Label-free enrichment of adrenal cortical progenitor cells using inertial microfluidics. PloS one, 7(10):e46550, January 2012. [44] Soojung Claire Hur, Sung-Eun Choi, Sunghoon Kwon, and Dino Di Carlo. Inertial focusing of non-spherical microparticles. Applied Physics Letters, 99(4):044101, 2011. [45] Soojung Claire Hur, Nicole K. Henderson-MacLennan, Edward R. B. McCabe, and Dino Di Carlo. Deformability-based cell classification and enrichment using inertial microfluidics. Lab on a Chip, (11):912–920, 2011. [46] Hong Miao Ji, Victor Samper, Yu Chen, Chew Kiat Heng, Tit Meng Lim, and Levent Yobas. Silicon-based microfilters for whole blood cell separation. Biomedical microdevices, 10(2):251–7, April 2008. [47] D D Joseph, D Ocando, and P Y Huang. Slip velocity and lift. Journal of Fluid Mechanics, 454:263–286, 2002. [48] D Kim, E Choi, S S Choi, S Lee, J Park, and K Yun. Measurement of Single-Cell Deformability Using Impedance Analysis on Microfluidic Chip. Japanese Journal of Applied Physics, 49(12):127002, December 2010. [49] Sathyakumar S Kuntaegowdanahalli, Ali Asgar S Bhagat, Girish Kumar, and Ian Papautsky. Inertial microfluidics for continuous particle separation in spiral microchannels. Lab on a chip, 9(20):2973–80, October 2009. [50] Ryoichi Kurose and Satoru Komori. Drag and lift forces on a rotating sphere in a linear shear flow. Journal of Fluid Mechanics, 384:183–206, April 1999. 108 REFERENCES [51] Kiho Kwon, TaeSeok Sim, Hui-Sung Moon, Jeong-Gun Lee, Jae Chan Park, and Hyo-Il Jung. a Novel Particle Separation Method using MultiStage Multi-Orifice Flow Fractionation(MS-MOFF). In microTAS, number October, pages 199–201, 2010. [52] Wilbur A Lam, Michael J Rosenbluth, and Daniel A Fletcher. Chemotherapy exposure increases leukemia cell stiffness. Blood, 109(8):3505–3508, 2007. [53] Franziska Lautenschl¨ager, Stephan Paschke, Stefan Schinkinger, Arlette Bruel, Michael Beil, and Jochen Guck. The regulatory role of cell mechanics for migration of differentiating myeloid cells. Proceedings of the National Academy of Sciences of the United States of America, 106(37):15696–701, September 2009. [54] Myung Gwon Lee, Chae Yun Bae, Sungyoung Choi, Hyun-jung Cho, and Je-kyun Park. Hight-Throughput Inertial Separation Of Cancer Cells From Human Whole Blood In A Contraction-Expansion Array Microchannel. In microTAS, pages 2065–2067, 2011. [55] Myung Gwon Lee, Sungyoung Choi, and Je-Kyun Park. Inertial separation in a contraction-expansion array microchannel. Journal of chromatography. A, 1218(27):4138–43, July 2011. [56] Sung S Lee, Yoonjae Yim, Kyung H Ahn, and Seung J Lee. Extensional flow-based assessment of red blood cell deformability using hyperbolic converging microchannel. Biomedical microdevices, May 2009. [57] Wong Cheng Lee, Ali Asgar S Bhagat, Sha Huang, Krystyn J Van Vliet, and Jongyoon Han. Cell Cycle Synchronization of Stem Cells Using Inertial Microfluidics. In microTAS, number October, pages 208–210, 2010. [58] Wong Cheng Lee, Ali Asgar S Bhagat, Sha Huang, Krystyn J Van Vliet, Jongyoon Han, and Chwee Teck Lim. High-throughput cell cycle synchronization using inertial forces in spiral microchannels. Lab on a chip, 11(7):1359–67, April 2011. [59] M Lekka, P Laidler, D Gil, J Lekki, Z Stachura, and a Z Hrynkiewicz. Elasticity of normal and cancerous human bladder cells studied by scanning force microscopy. European biophysics journal : EBJ, 28(4):312–6, January 1999. [60] Bryan Lincoln, Harold M Erickson, Stefan Schinkinger, Falk Wottawah, Daniel Mitchell, Sydney Ulvick, Curt Bilby, and Jochen Guck. 109 REFERENCES Deformability-based flow cytometry. 2004. Cytometry A, 59(2):203–9, June [61] Bryan Lincoln, Stefan Schinkinger, Kort Travis, Falk Wottawah, Susanne Ebert, Frank Sauer, and Jochen Guck. Reconfigurable microfluidic integration of a dual-beam laser trap with biomedical applications. Biomedical microdevices, 9(5):703–10, October 2007. [62] Wei-Ming Lu, Kuo-Lun Tung, Shu-Mei Hung, Jia-Shyan Shiau, and KuoJen Hwang. Compression of deformable gel particles. Powder Technology, 116(1):1–12, May 2001. [63] M P MacDonald, G C Spalding, and K Dholakia. Microfluidic sorting in an optical lattice. Nature, 426:421–424, 2003. [64] Albert J Mach and Dino Di Carlo. Continuous scalable blood filtration device using inertial microfluidics. Biotechnology and bioengineering, 107(2):302–11, October 2010. [65] Albert J Mach, Jae Hyun Kim, Armin Arshi, Soojung Claire Hur, and Dino Di Carlo. Automated cellular sample preparation using a Centrifuge-on-aChip. Lab on a chip, 11(17):2827–34, September 2011. [66] R E Mahaffy, C K Shih, F C MacKintosh, and J K¨as. Scanning probe-based frequency-dependent microrheology of polymer gels and biological cells. Physical review letters, 85(4):880–3, July 2000. [67] Joseph M Martel and Mehmet Toner. Inertial focusing dynamics in spiral microchannels. Physics of Fluids, 24(3):032001, 2012. [68] Hisham Mohamed, Megan Murray, James N Turner, and Michele Caggana. Isolation of tumor cells using size and deformation. Journal of chromatography. A, 1216(47):8289–95, November 2009. [69] S A Morsi and A J Alexander. An investigation of particle trajectories in two-phase flow systems. Journal of Fluid Mechanics, 55(02):193–208, March 2006. [70] Keith J Morton, Kevin Loutherback, David W Inglis, Ophelia K Tsui, James C Sturm, Stephen Y Chou, and Robert H Austin. Crossing microfluidic streamlines to lyse, label and wash cells. Lab on a chip, 8(9):1448–53, September 2008. 110 REFERENCES [71] Keith J Morton, Kevin Loutherback, David W Inglis, Ophelia K Tsui, James C Sturm, Stephen Y Chou, and Robert H Austin. Hydrodynamic metamaterials: microfabricated arrays to steer, refract, and focus streams of biomaterials. Proceedings of the National Academy of Sciences of the United States of America, 105(21):7434–8, May 2008. [72] Jeonghun Nam, Yongjin Lee, and Sehyun Shin. Size-dependent microparticles separation through standing surface acoustic waves. Microfluidics and Nanofluidics, 11(3):317–326, April 2011. [73] Beyer N Nardi and da Silva L Meirelles. Mesenchymal stem cells: isolation, in vitro expansion and characterization. Handbook of experimental pharmacology, 174:249–82, January 2006. [74] Harm A Nieuwstadt, Robinson Seda, David S Li, J Brian Fowlkes, and Joseph L Bull. Microfluidic particle sorting utilizing inertial lift force. Biomedical microdevices, 13(1):97–105, February 2011. [75] Pandharinath S Nikam, Meera C Jadhav, Mehdi Hasan, Physical Chemistry, M S G College, and Malegaon Camp. Density and Viscosity of Mixtures of Dimethyl Sulfoxide + Methanol, +Ethanol, +Propan-1-ol, +Propan-2-ol, +Butan-1-ol, +2-Methylpropan-1-ol, and +2-Methylpropan-2-ol at 298.15 K and 303.15 K. Journal of Chemical & Engineering Data, 2(1):1028– 1031, 1996. [76] N Nivedita, P V Giridhar, S Kasper, and I Papautsky. Sorting Human Prostate Epithelial(HPET) Cells In An Inertial Microfluidic Device. In microTAS, number c, pages 1230–1232, 2011. [77] Shinichi Ookawara, Ryochi Higashi, David Street, and Kohei Ogawa. Feasibility study on concentration of slurry and classification of contained particles by microchannel. Chemical Engineering Journal, 101(1-3):171–178, August 2004. [78] Shinichi Ookawara, Nobuo Oozeki, Kohei Ogawa, Patrick L¨ob, and Volker Hessel. Process intensification of particle separation by lift force in arc microchannel with bifurcation. Chemical Engineering and Processing: Process Intensification, 49(7):697–703, July 2010. [79] Shinichi Ookawara, David Street, and Kohei Ogawa. Numerical study on development of particle concentration profiles in a curved microchannel. Chemical Engineering Science, 61(11):3714–3724, June 2006. 111 REFERENCES [80] World Health Organization. Cancer. http://www.who.int/ mediacentre/factsheets/fs297/en/, February 2009. [81] Nicole Pamme and Andreas Manz. On-chip free-flow magnetophoresis: continuous flow separation of magnetic particles and agglomerates. Analytical chemistry, 76(24):7250–6, December 2004. [82] NA Patankar and PY Huang. Lift-off of a single particle in Newtonian and viscoelastic fluids by direct numerical simulation. Journal of Fluid . . . , 438:67–100, 2001. [83] Filip Petersson, Lena Aberg, Ann-Margret Sw¨ard-Nilsson, and Thomas Laurell. Free flow acoustophoresis: microfluidic-based mode of particle and cell separation. Analytical chemistry, 79(14):5117–23, July 2007. [84] C Prohm, M Gierlak, and H Stark. Inertial microfluidics with multiparticle collision dynamics. The European physical journal. E, Soft matter, 35(8):9757, August 2012. [85] Michael J Rosenbluth, Wilbur a Lam, and Daniel a Fletcher. Force microscopy of nonadherent cells: a comparison of leukemia cell deformability. Biophysical journal, 90(8):2994–3003, April 2006. [86] Christian Rotsch and Manfred Radmacher. Drug-Induced Changes of Cytoskeletal Structure and Mechanics in Fibroblasts: An Atomic Force Microscopy Study. Biophysical Journal, 78(1):520–535, January 2000. [87] Aman Russom, Amit K Gupta, Sunitha Nagrath, Dino Di Carlo, Jon F Edd, and Mehmet Toner. Differential inertial focusing of particles in curved lowaspect-ratio microchannels. New journal of physics, 11:75025, July 2009. [88] P G Saffman. The lift on a small sphere in a slow shear flow. Journal of Fluid Mechanics, 22(02):385–400, March 1965. [89] Jeffrey A Schongerg and E J Hinch. Inertial migration of a sphere in Poiseuille flow. Journal of Fluid Mechanics, 203:517–525, December 1989. [90] Jeonggi Seo, Meng H. Lean, and Ashutosh Kole. Membrane-free microfiltration by asymmetric inertial migration. Applied Physics Letters, 91(3):033901, 2007. [91] Jeonggi Seo, Meng H Lean, and Ashutosh Kole. Membraneless microseparation by asymmetry in curvilinear laminar flows. Journal of chromatography. A, 1162(2):126–31, August 2007. 112 REFERENCES [92] G. J. Sheard, K. Hourigan, and M. C. Thompson. Computations of the drag coefficients for low-Reynolds-number flow past rings. Journal of Fluid Mechanics, 526:257–275, March 2005. [93] J Patrick Shelby, John White, Karthikeyan Ganesan, Pradipsinh K Rathod, and Daniel T Chiu. A microfluidic model for single-cell capillary obstruction by Plasmodium falciparum- infected erythrocytes. PNAS, 100(25):14618–14622, 2003. [94] John Sleep, David Wilson, Robert Simmons, and Walter Gratzer. Elasticity of the red cell membrane and its relation to hemolytic disorders: an optical tweezers study. Biophysical Journal, 77(6):3085–95, December 1999. [95] Jiashu Sun, Mengmeng Li, Chao Liu, Yi Zhang, Dingbin Liu, Wenwen Liu, Guoqing Hu, and Xingyu Jiang. Double spiral microchannel for labelfree tumor cell separation and enrichment. Lab on a chip, 12(20):3952–60, September 2012. [96] Subra Suresh. Biomechanics and biophysics of cancer cells. Acta biomaterialia, 3(4):413–38, July 2007. [97] Swee Jin Tan, Rumkumar Lalitha Lakshmi, Pengfei Chen, Wan-Teck Lim, Levent Yobas, and Chwee Teck Lim. Versatile label free biochip for the detection of circulating tumor cells from peripheral blood in cancer patients. Biosensors & bioelectronics, 26(4):1701–5, December 2010. [98] Swee Jin Tan, Levent Yobas, Gabriel Yew Hoe Lee, Choon Nam Ong, and Chwee Teck Lim. Microdevice for the isolation and enumeration of cancer cells from blood. Biomedical microdevices, 11(4):883–92, August 2009. [99] Tatsuya Tanaka, Takuji Ishikawa, Keiko Numayama-Tsuruta, Yohsuke Imai, Hironori Ueno, Takefumi Yoshimoto, Noriaki Matsuki, and Takami Yamaguchi. Inertial migration of cancer cells in blood flow in microchannels. Biomedical microdevices, 14(1):25–33, February 2012. [100] Abel L Thangawng, Rodney S Ruoff, Melody a Swartz, and Matthew R Glucksberg. An ultra-thin PDMS membrane as a bio/micro-nano interface: fabrication and characterization. Biomedical Microdevices, 9(4):587–95, August 2007. [101] Michael D Vahey and Joel Voldman. An equilibrium method for continuous-flow cell sorting using dielectrophoresis. Analytical chemistry, 80(9):3135–43, May 2008. 113 REFERENCES [102] Siva a Vanapalli, Michel H G Duits, and Frieder Mugele. Microfluidics as a functional tool for cell mechanics. Biomicrofluidics, 3(1):12006, January 2009. [103] D J Vojir and E E Michaelides. Effect of the history term on the motion of rigid spheres in a viscous fluid. International journal of multiphase flow, 20(3):547–556, 1994. [104] Mark M Wang, Eugene Tu, Daniel E Raymond, Joon Mo Yang, Haichuan Zhang, Norbert Hagen, Bob Dees, Elinore M Mercer, Anita H Forster, Ilona Kariv, Philippe J Marchand, and William F Butler. Microfluidic sorting of mammalian cells by optical force switching. Nature biotechnology, 23(1):83–7, January 2005. [105] Xiao Wang, Jian Zhou, and Ian Papautsky. Continuous Rare Cell Extraction Using Self-releasing Vortex in an Inertial Microfluidic Device. In microTAS, number c, pages 533–535, 2012. [106] Huibin Wei, Bor-han Chueh, Huiling Wu, Eric W Hall, Cheuk-wing Li, Romana Schirhagl, Jin-Ming Lin, and Richard N Zare. Particle sorting using a porous membrane in a microfluidic device. Lab on a chip, 11(2):238–45, January 2011. [107] Peter Wilding, Larry J Kricka, Jing Cheng, Gia Hvichia, Mann a Shoffner, and Paolo Fortina. Integrated cell isolation and polymerase chain reaction analysis using silicon microfilter chambers. Analytical biochemistry, 257(2):95–100, March 1998. [108] Lidan Wu, Guofeng Guan, Han Wei Hou, Ali Asgar S Bhagat, and Jongyoon Han. Separation of leukocytes from blood using spiral channel with trapezoid cross-section. Analytical chemistry, 84(21):9324–31, November 2012. [109] Zhigang Wu, Ben Willing, Joakim Bjerketorp, Janet K Jansson, and Klas Hjort. Soft inertial microfluidics for high throughput separation of bacteria from human blood cells. Lab on a chip, 9(9):1193–9, May 2009. [110] Bernhard K Wunderlich, Ulrich a Klessinger, and Andreas R Bausch. Diffusive spreading of time-dependent pressures in elastic microfluidic devices. Lab on a chip, 10(8):1025–9, April 2010. [111] Masumi Yamada, Megumi Nakashima, and Minoru Seki. Pinched flow fractionation: continuous size separation of particles utilizing a laminar 114 REFERENCES flow profile in a pinched microchannel. Analytical chemistry, 76(18):5465– 71, September 2004. [112] Masumi Yamada and Minoru Seki. Hydrodynamic filtration for on-chip particle concentration and classification utilizing microfluidics. Lab on a chip, 5(11):1233–9, November 2005. [113] B H Yang, J Wang, D D Joseph, Howard H Hu, T-W Pan, and R Glowinski. Migration of a sphere in tube flow. Journal of Fluid Mechanics, 540:109– 131, 2005. [114] Siyang Zheng, Henry Lin, Jing-Quan Liu, Marija Balic, Ram Datar, Richard J Cote, and Yu-Chong Tai. Membrane microfilter device for selective capture, electrolysis and genomic analysis of human circulating tumor cells. Journal of chromatography. A, 1162(2):154–61, August 2007. [115] Siyang Zheng, Yu-Chong Tai, and Harvey Kasdan. A micro device for separation of erythrocytes and leukocytes in human blood. In Annual International Conference of the IEEE Engineering in Medicine and Biology Society., volume 1, pages 1024–7, January 2005. [116] Jian Zhou, P V Giridhar, S Kasper, and Ian Papautsky. Extraction and Enrichment of Rare Cells in a Simple Inertial Microfluidic Device. In microTAS, number c, pages 1906–1908, 2011. [117] Junjie Zhu, Robert Cameron Canter, Gyunay Keten, Pallavi Vedantam, Tzuen-Rong J. Tzeng, and Xiangchun Xuan. Continuous-flow particle and cell separations in a serpentine microchannel via curvature-induced dielectrophoresis. Microfluidics and Nanofluidics, 11(6):743–752, July 2011. 115 [...]... Technology for Lab-On-a-Chip, Size and Deformability Based Cell Sorting There are several emerging lab-on-a-chip microfluidic technologies in label free cell sorting, especially based on cell size or deformability In size based sorting, there are positive sorting that applies external optical/magnetic/electrical force fields across the flow stream of cell flow stream and negative sorting methods that only rely on... can be classified as either size or deformability based sorting Thus, in the following sections, size and deformability based methods are discussed separately 2.1 Microfluidics Methodologies for Size Based Cell Separation Size is one of the basic physical properties of a cell Many micro-scale cell separation techniques take advantage of this intrinsic property for high performance separation These techniques... advantages over conventional methods, their performance (in terms of throughput, efficiency, and robustness) remains low This hinders their usability in clinical applications 5 Introduction 1.3 Objective of the New Size and Deformability Based Cell Sorting Methods Development There exist several research gaps for current study on size and deformability based cell sorting and separation Although inertial microfluidics... hydrodynamic forces generated in the flow, such as drag force and inertial lift force Several corresponding technologies, such as external optical force field, mechanical pressing, and hydrodynamic stretching were also used for deformability based sorting These merging technologies make separation and sorting of many types of cells possible, especially for those cells for which proper antibody for labelling... cell sorting and separation methods and the theory underlying In the theoretical aspect, the cell/ particle focusing mechanism in inertial microfluidic with curved channel is expected to be revealed in size based sorting A model of squeezing process of viscoelastic cell through a narrow gap will be constructed for deformability based sorting In the application aspect, a novel technique for size based cell. .. properties of sorted cells Chapter 6 summarises the work that has been done in this thesis and outlines some future research directions 7 Chapter 2 Background and Literature Review In this chapter, methodologists on size and deformability based sorting and measuring in single cell level are reviewed Although there are some crossover technologies that considered both size and deformability in sorting, most... healthy cells based on their stiffness difference Similar to the case of cancerous and healthy cells, malaria infected red blood cells are stiffer than normal red blood cells, and diagnosis methods based on stiffness difference have been demonstrated These examples accentuate the need for, and the potential of research in label-free separation and sorting Although existing label-free cell separation and sorting. .. MSCs differentiate to different cell types, both the cell size and deformability will change Size- and deformability- based sorting devices constitute good methodology to study the bioproperties during this differentiation process 2 Introduction 1.1.2 Cancer and Circulating Tumor Cells Cancer is one of the leading causes of death worldwide Cancer affects people at all ages and the risk increases with age... conditions and derive the forces subjected on the particle (ii) Develop an active controlled microfluidic channel with optical feedback of size information to gauge cell deformability considering size variation (iii) Build a mathematical model for the deformability based sorting device to evaluate the stiffness of measured cell populations The results of this study may have significant impact on label-free cell. .. efficiency size based separation chip in Chapter 4 Chapter 4 presents a microfluidic spiral channel with trapezoidal cross section The particle focusing and trapping behaviour are studied The separation performance of rigid beads, blood cells, and human mesenchymal stem cells are given Chapter 5 presents the development of deformability based cell sorting technology Sorting data of several types of cells . Requiremnts of New Technology for Lab-On-a-Chip, Size and Deformability Based Cell Sorting . . . . . . . . . . . . . . . 5 1.3 Objective of the New Size and Deformability Based Cell Sorting Methods Development. technologies, new de- vices for size- and deformability- based cell sorting were designed and their per- formance studied. Firstly, for developing high throughput size- based separation method, three- dimensional. MICROFLUIDICS FOR SIZE AND DEFORMABILITY BASED CELL SORTING GUAN GUOFENG A THESIS SUBMITTED FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT