Drag controlled formation of polymeric colloids with optical traps

Erel Lasnoy, Omer Wagner, Eitan Edri and Hagay Shpaisman

Optical trapping is a powerful optical manipulation technique for controlling various mesoscopic systems that allows formation of tailor-made polymeric micro-sized colloids by directed coalescence of nucleation sites. However, control over the size of a single colloid requires constant monitoring of the growth process and deactivation of the optical trap once it reaches the required dimensions. Moreover, producing more than one colloid requires moving the sample to a pristine location where the process must be repeated. Here, we present a novel method for continuous control over formation of polydimethylsiloxane colloids based on directed coalescence induced by optical traps under flow inside microfluidic channels. Once the drag force on a growing colloid exceeds the trapping force, it leaves the optical trap, and a new colloid starts to form at the same location. We demonstrate repeatability of the process and selectively produce colloids with radii of ∼1–14 μm by controlling the laser intensity and flow rate. In addition, holographic optical tweezers are used to show how multiple optical traps in 3D could be used to influence a significant cross section of the micro-channel, thus forming a light-controlled assembly line for colloidal formation.

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Generating digital drug cocktails via optical manipulation of drug-containing particles and photo-patterning of hydrogels

Yi-Sin Chen, Ko-Chin Chung, Wen-Yen Huang, Wen-Bin Lee, Chien-Yu Fu, Chih-Hung Wang and Gwo-Bin Lee

An integrated microfluidic system combining 1) an optically-induced-dielectrophoresis (ODEP) module for manipulation of drug-containing particles and 2) an ultraviolet (UV) “direct writing” module capable of patterning hydrogels was established herein for automatic formulation of customized digital drug cocktails. Using the ODEP module, the drug-containing particles were assembled by using moving light patterns generated from a digital projector. The hydrogel, poly(ethylene glycol) diacrylate (PEGDA), was used as the medium in the ODEP module such that the assembled drug-containing particles could be UV-cured and consequently encapsulated in “pills” of specific sizes and shapes by using the UV direct writing module. At an optimal ODEP force of 335 pN, which was achieved in a solution of 15% PEGDA in 0.2 M sucrose, it was possible to manipulate and UV-cure the drug-containing particles. Furthermore, with a digital micromirror device inside the UV direct writing module, different UV patterns could be designed and projected, allowing for the digital drug cocktails to be packaged into different shapes in <60 s. As a demonstration, emulsion droplets containing two different anti-cancer drugs were further tested to show the capability of the developed device. This represents an automatic digital drug cocktail formulating device which stands to revolutionize personalized medicine.

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Quantitative phase microscopy of red blood cells during planar trapping and propulsion

Azeem Ahmad, Vishesh Dubey, Vijay Raj Singh, Jean-Claude Tinguely, Cristina Ionica Øie, Deanna L. Wolfson, Dalip Singh Mehta, Peter T. C. So and Balpreet Singh Ahluwalia

Red blood cells (RBCs) have the ability to undergo morphological deformations during microcirculation, such as changes in surface area, volume and sphericity. Optical waveguide trapping is suitable for trapping, propelling and deforming large cell populations along the length of the waveguide. Bright field microscopy employed with waveguide trapping does not provide quantitative information about structural changes. Here, we have combined quantitative phase microscopy and waveguide trapping techniques to study changes in RBC morphology during planar trapping and transportation. By using interference microscopy, time-lapsed interferometric images of trapped RBCs were recorded in real-time and subsequently utilized to reconstruct optical phase maps. Quantification of the phase differences before and after trapping enabled study of the mechanical effects during planar trapping. During planar trapping, a decrease in the maximum phase values, an increase in the surface area and a decrease in the volume and sphericity of RBCs were observed. QPM was used to analyze the phase values for two specific regions within RBCs: the annular rim and the central donut. The phase value of the annular rim decreases whereas it increases for the central donut during planar trapping. These changes correspond to a redistribution of cytosol inside the RBC during planar trapping and transportation.

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Optical tweezing using tunable optical lattices along a few-mode silicon waveguide

C. Pin, J.-B. Jager, M. Tardif, E. Picard, E. Hadji, F. de Fornel and B. Cluzel

Fourteen years ago, optical lattices and holographic tweezers were considered as a revolution, allowing for trapping and manipulating multiple particles at the same time using laser light. Since then, near-field optical forces have aroused tremendous interest as they enable efficient trapping of a wide range of objects, from living cells to atoms, in integrated devices. Yet, handling at will multiple objects using a guided light beam remains a challenging task for current on-chip optical trapping techniques. We demonstrate here on-chip optical trapping of dielectric microbeads and bacteria using one-dimensional optical lattices created by near-field mode beating along a few-mode silicon nanophotonic waveguide. This approach allows not only for trapping large numbers of particles in periodic trap arrays with various geometries, but also for manipulating them via diverse transport and repositioning techniques. Near-field mode-beating optical lattices may be readily implemented in lab-on-a-chip devices, addressing numerous scientific fields ranging from bio-analysis to nanoparticle processing.

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Measurement of the mechanical properties of single Synechocystis sp. strain PCC6803 cells in different osmotic concentrations using a robot-integrated microfluidic chip

Di Chang, Shinya Sakuma, Kota Kera, Nobuyuki Uozumi and Fumihito Arai

Synechocystis sp. strain PCC6803 (Synechocystis) is a model microorganism and its mechanosensitive (MS) channels play important roles in its osmoadaptation mechanism. When the osmotic concentration of the culture environment changes, the inner pressure of the cell also changes due to the transportation of water through ion channels. Because the tension in the cell membrane relates to the inner pressure, we expect that the response of the MS channels to an osmotic concentration change could be evaluated by measuring their mechanical properties. Here, we propose a system for the measurement of the mechanical properties of a single Synechocystis cell. We developed a robot-integrated microfluidic chip combined with optical tweezers. The chip has an external actuated pushing probe and a force sensor probe. A single cell was located between the tip of both probes using the optical tweezers and was then deformed using the probes. As a result, we could measure the force and deformation and compare the Young's moduli of two groups: a group of wild type cells and a group of mutant (genetically modified) cells with a defect in the MS channels, at three different osmotic concentrations. The results showed that the Young's modulus of each group changed according to the osmotic concentration, while changes in cell size were too small to be detected. These results confirmed that the proposed evaluation method provides an understanding of the physiological function of MS channels for keeping the cell integrity of microorganisms when the cells are exposed to different external osmotic changes.

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Impact of complex surfaces on biomicrorheological measurements using optical tweezers

Shu Zhang, Lachlan J. Gibson, Alexander B. Stilgoe, Timo A. Nieminen and Halina Rubinsztein-Dunlop

The characterisation of physical properties in biologically relevant processes and the development of novel microfluidic devices for this purpose are experiencing a great resurgence at present. In many of measurements of this type where a probe in a fluid is used, the strong influence of the boundaries of the volume used is a serious problem. In these geometries the proximity of a probe to a wall can severely influence the measurement. However, although much knowledge has been gained about flat walls, to date, the effect of non-planar surfaces at microscopic scale on rotational motion of micro-objects has not been studied. Here we present for the first time both experimental measurements and numerical computations which aim to study the drag torque on optically trapped rotating particles moving near 3D-printed conical and cylindrical walls on-chip. These results are essential for quantifying how curved walls can effect the torque on particles, and thus enable accurate hydrodynamic simulations at the micron-scale. This opens the potential for new sensing approaches under more complex conditions, allowing both dynamic and microrheological studies of biological systems and lab-on-chip devices.

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High-resolution and multi-range particle separation by microscopic vibration in an optofluidic chip

Y. Z. Shi, S. Xiong, L. K. Chin, Y. Yang, J. B. Zhang, W. Ser, J. H. Wu, T. N. Chen, Z. C. Yang, Y. L. Hao, B. Liedberg, P. H. Yap, Y. Zhang and A. Q. Liu

An optofluidic chip is demonstrated in experiments for high-resolution and multi-range particle separation through the optically-induced microscopic vibration effect, where nanoparticles are trapped in loosely overdamped optical potential wells created with combined optical and fluidic constraints. It is the first demonstration of separating single nanoparticles with diameters ranging from 60 to 100 nm with a resolution of 10 nm. Nanoparticles vibrate with an amplitude of 3–7 μm in the loosely overdamped potential wells in the microchannel. The proposed optofluidic device is capable of high-resolution particle separation at both nanoscale and microscale without reconfiguring the device. The separation of bacteria from other larger cells is accomplished using the same chip and operation conditions. The unique trapping mechanism and the superb performance in high-resolution and multi-range particle separation of the proposed optofluidic chip promise great potential for a diverse range of biomedical applications.

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Microfluidic-based high-throughput optical trapping of nanoparticles

Abhay Kotnala, Yi Zheng, Jianping Fu and Wei Cheng
Optical tweezers have emerged as a powerful tool for multiparametric analysis of individual nanoparticles with single-molecule sensitivity. However, its inherent low-throughput characteristic remains a major obstacle to its applications within and beyond the laboratory. This limitation is further exacerbated when working with low concentration nanoparticle samples. Here, we present a microfluidic-based optical tweezers system that can ‘actively’ deliver nanoparticles to a designated microfluidic region for optical trapping and analysis. The active microfluidic delivery of nanoparticles results in significantly improved throughput and efficiency for optical trapping of nanoparticles. We observed a more than tenfold increase in optical trapping throughput for nanoparticles as compared to conventional systems at the same nanoparticle concentration. To demonstrate the utility of this microfluidic-based optical tweezers system, we further used back-focal plane interferometry coupled with a trapping laser for the precise quantitation of nanoparticle size without prior knowledge of the refractive index of nanoparticles. The development of this microfluidic-based active optical tweezers system thus opens the door to high-throughput multiparametric analysis of nanoparticles using precision optical traps in the future.

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A liquid thermal gradient refractive index lens and using it to trap single living cell in flowing environments

H. L. Liu, Y. Shi, L. Liang, L. Li, S. S. Guo, L. Yin and Y. Yang

A gradient refractive index (GRIN) lens has a great potential for on-chip imaging and detection systems because of its flat surface with reduced defects. This paper reports a liquid thermal GRIN lens prepared using heat conduction between only one liquid, and uses it as a tunable optical tweezer for single living cell trapping in a flowing environment. This liquid GRIN lens consists of a trapezoidal region in the upper layer which is used to establish a GRIN profile by the heat conduction between three streams of benzyl alcohol with different temperatures, and subsequently a rhombus region in the lower layer with compensation liquids to form a steady square-law parabolic refractive index profile only in transverse direction. Simulations and experiments successfully show the real-time tunability of the focusing properties. The focal length can be modulated in the range of 500 μm with the minimum focal length of 430 μm. A considerable high enhancement factor achieves 5.4 whereas the full width at half maximum is 4 μm. The response time of the GRIN lens is about 20 ms. Based on this enhancement, tunable optical trapping for single human embryonic kidney 293 cell in the range of 280 μm is demonstrated by varying the focal length and working distance which is difficult for solid optical tweezers. The considerable quality of this liquid GRIN lens indicates on-chip applications especially in high quality optical imaging, detection and cells' handling.

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Onset of particle trapping and release via acoustic bubbles

Yun Chen, Zecong Fang, Brett Merritt, Dillon Strack, Jie Xu and Sungyon Lee

Trapping and sorting of micro-sized objects is one important application of lab on a chip devices, with the use of acoustic bubbles emerging as an effective, non-contact method. Acoustically actuated bubbles are known to exert a secondary radiation force (FSR) on micro-particles and stabilize them on the bubble surface, when this radiation force exceeds the external hydrodynamic forces that act to keep the particles in motion. While the theoretical expression of FSR has been derived by Nyborg decades ago, no direct experimental validation of this force has been performed, and the relationship between FSR and the bubble's ability to trap particles in a given lab on a chip device remains largely empirical. In order to quantify the connection between the bubble oscillation and the resultant FSR, we experimentally measure the amplitude of bubble oscillations that give rise to FSR and observe the trapping and release of a single microsphere in the presence of the mean flow at the corresponding acoustic parameters using an acoustofluidic device. By combining well-developed theories that connect bubble oscillations to the acoustic actuation, we derive the expression for the critical input voltage that leads to particle release into the flow, in good agreement with the experiments.

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Cell stretching devices as research tools: engineering and biological considerations

Harshad Kamble, Matthew J. Barton, Myeongjun Jun, Sungsu Park and Nam-Trung Nguyen

Cells within the human body are subjected to continuous, cyclic mechanical strain caused by various organ functions, movement, and growth. Cells are well known to have the ability to sense and respond to mechanical stimuli. This process is referred to as mechanotransduction. A better understanding of mechanotransduction is of great interest to clinicians and scientists alike to improve clinical diagnosis and understanding of medical pathology. However, the complexity involved in in vivo biological systems creates a need for better in vitro technologies, which can closely mimic the cells' microenvironment using induced mechanical strain. This technology gap motivates the development of cell stretching devices for better understanding of the cell response to mechanical stimuli. This review focuses on the engineering and biological considerations for the development of such cell stretching devices. The paper discusses different types of stretching concepts, major design consideration and biological aspects of cell stretching and provides a perspective for future development in this research area.

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Imaging the position-dependent 3D force on micro beads subjected to acoustic radiation forces and streaming

Andreas Lamprecht, Stefan Lakämper, Thierry Baasch, Iwan A.T. Schaap and Jürg Dual

Acoustic particle manipulation in microfluidic channels is becoming a powerful tool in microfluidics to control micrometer sized objects in medical, chemical and biological applications. By creating a standing acoustic wave in the channel, the resulting pressure field can be employed to trap or sort particles. To design efficient and reproducible devices, it is important to characterize the pressure field throughout the volume of the microfluidic device. Here, we used an optically trapped particle as probe to measure the forces in all three dimensions. By moving the probe through the volume of the channel, we imaged spatial variations in the pressure field. In the direction of the standing wave this revealed a periodic energy landscape for 2 μm beads, resulting in an effective stiffness of 2.6 nN/m for the acoustic trap. We found that multiple fabricated devices showed consistent pressure fields. Surprisingly, forces perpendicular to the direction of the standing wave reached values of up to 20% of the main-axis-values. To separate the direct acoustic force from secondary effects, we performed experiments with different bead sizes, which attributed some of the perpendicular forces to acoustic streaming. This method to image acoustically generated forces in 3D can be used to either minimize perpendicular forces or to employ them for specific applications in novel acoustofluidic designs.

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Acoustic force mapping in a hybrid acoustic-optical micromanipulation device supporting high resolution optical imaging

Gregor Thalhammer, Craig McDougall, Mike P MacDonald and Monika Ritsch-Marte

Many applications in the life-sciences demand non-contact manipulation tools for forceful but nevertheless delicate handling of various types of sample. Moreover, the system should support high-resolution optical imaging. Here we present a hybrid acoustic/optical manipulation system which utilizes a transparent transducer, making it compatible with high-NA imaging in a microfluidic environment. The powerful acoustic trapping within a layered resonator, which is suitable for highly parallel particle handling, is complemented by the flexibility and selectivity of holographic optical tweezers, with the specimens being under high quality optical monitoring at all times. The dual acoustic/optical nature of the system lends itself to optically measure the exact acoustic force map, by means of direct force measurements on an optically trapped particle. For applications with (ultra-)high demand on the precision of the force measurements, the position of the objective used for the high-NA imaging may have significant influence on the acoustic force map in the probe chamber. We have characterized this influence experimentally and the findings were confirmed by model simulations. We show that it is possible to design the chamber and to choose the operating point in such a way as to avoid perturbations due to the objective lens. Moreover, we found that measuring the electrical impedance of the transducer provides an easy indicator for the acoustic resonances.

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An inverted dielectrophoretic device for analysis of attached single cell mechanics

Rebecca Lownes Urbano and Alisa Morss Clyne

Dielectrophoresis (DEP), the force induced on a polarizable body by a non-uniform electric field, has been widely used to manipulate single cells in suspension and analyze their stiffness. However, most cell types do not naturally exist in suspension but instead require attachment to the tissue extracellular matrix in vivo. Cells alter their cytoskeletal structure when they attach to a substrate, which impacts cell stiffness. It is therefore critical to be able to measure mechanical properties of cells attached to a substrate. We present a novel inverted quadrupole dielectrophoretic device capable of measuring changes in the mechanics of single cells attached to a micropatterned polyacrylamide gel. The device is positioned over a cell of defined size, a directed DEP pushing force is applied, and cell centroid displacement is dynamically measured by optical microscopy. Using this device, single endothelial cells showed greater centroid displacement in response to applied DEP pushing force following actin cytoskeleton disruption by cytochalasin D. In addition, transformed mammary epithelial cell (MCF10A-NeuT) showed greater centroid displacement in response to applied DEP pushing force compared to untransformed cells (MCF10A). DEP device measurements were confirmed by showing that the cells with greater centroid displacement also had a lower elastic modulus by atomic force microscopy. The current study demonstrates that an inverted DEP device can determine changes in single attached cell mechanics on varied substrates.

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Trapping and viability of swimming bacteria in an optoelectric trap

A. Mishra, T. R. Maltais, T. M. Walter, A. Wei, S. J. Williams and S. T. Wereley

Non-contact manipulation methods capable of trapping and transporting swimming bacteria can significantly aid in chemotaxis studies. However, high swimming speed makes the trapping of these organisms an inherently challenging task. We demonstrate that an optoelectric technique, rapid electrokinetic patterning (REP), can effectively trap and manipulate Enterobacter aerogenes bacteria swimming at velocities greater than 20 μm s−1. REP uses electro-orientation, laser-induced AC electrothermal flow, and particle–electrode interactions for capturing these cells. In contrast to trapping non-swimming bacteria and inert microspheres, we observe that electro-orientation is critical to the trapping of the swimming cells, since unaligned bacteria can swim faster than the radially inward electrothermal flow and escape the trap. By assessing the cell membrane integrity, we study the effect of REP trapping conditions, including optical radiation, laser-induced heating, and the electric field on cell viability. When applied individually, the optical radiation and laser-induced heating have negligible effect on cells. At the standard REP trapping conditions fewer than 2% of cells have a compromised membrane after four minutes. To our knowledge this is the first study detailing the effect of REP trapping on cell viability. The presented results provide a clear guideline on selecting suitable REP parameters for trapping living bacteria.

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Hybrid PDMS/glass microfluidics for high resolution imaging and application to sub-wavelength particle trapping

Mario Tonin, Nicolas Descharmes and Romuald Houdré

We demonstrate the fabrication of a hybrid PDMS/glass microfluidic layer that can be placed on top of non-transparent samples and allows high-resolution optical microscopy through it. The layer mimics a glass coverslip to limit optical aberrations and can be applied on the sample without the use of permanent bonding. The bonding strength can withstand to hold up to 7 bars of injected pressure in the channel without leaking or breaking. We show that this process is compatible with multilayer soft lithography for the implementation of flexible valves. The benefits of this application is illustrated by optically trapping sub-wavelength particles and manipulate them around photonic nano-structures. Among others, we achieve close to diffraction limited imaging through the microfluidic assembly, full control on the flow with no dynamical deformations of the membrane and a 20-fold improvement on the stiffness of the trap at equivalent trapping power.

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Frequency Modulated Microrheology

Matthew Myles Shindel and Eric M Furst

Coupling analog frequency modulation (FM) to the driving stimulus in active microrheology measurements conducted with optical tweezers effectively parallelizes numerous single-frequency experiments. Consequently, frequency modulated microrheology (FMMR) can efficiently characterize the dynamic stress response of complex fluids over several frequency decades in a single experiment. The time required to complete an FMMR measurement scales with the lowest frequency probed, improving throughput over the serial frequency sweep approach. The ease of implementation, straight-forward data analysis and rapidity of FMMR offer particular utility toward applications such as characterization of non-equilibrium materials, automated microrheology instrumentation, high-throughput screening of biomaterials and (bio)pharmaceutical formulations, and in-situ monitoring of chemical and biochemical reaction processes.

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An integrated optofluidic device for single-cell sorting driven by mechanical properties

T. Yang, P. Paiè, G. Nava, F. Bragheri, R. Martinez Vazquez, P. Minzioni, M. Veglione, M. Di Tano, C. Mondello, R. Osellame and I. Cristiani

We present a novel optofluidic device for real-time sorting on the basis of cell mechanical properties, measured by optical stretching. The whole mechanism, based on optical forces, does not hamper the viability of the tested cells, which can be used for further analysis. The device effectiveness is demonstrated by extracting a sample population enriched with highly metastatic cells from a heterogeneous cell mixture.

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Microfluidic cell sorting: a review of the advances in the separation of cells from debulking to rare cell isolation

C. Wyatt Shields IV, Catherine D. Reyes and Gabriel P. López

Accurate and high throughput cell sorting is a critical enabling technology in molecular and cellular biology, biotechnology, and medicine. While conventional methods can provide high efficiency sorting in short timescales, advances in microfluidics have enabled the realization of miniaturized devices offering similar capabilities that exploit a variety of physical principles. We classify these technologies as either active or passive. Active systems generally use external fields (e.g., acoustic, electric, magnetic, and optical) to impose forces to displace cells for sorting, whereas passive systems use inertial forces, filters, and adhesion mechanisms to purify cell populations. Cell sorting on microchips provides numerous advantages over conventional methods by reducing the size of necessary equipment, eliminating potentially biohazardous aerosols, and simplifying the complex protocols commonly associated with cell sorting. Additionally, microchip devices are well suited for parallelization, enabling complete lab-on-a-chip devices for cellular isolation, analysis, and experimental processing. In this review, we examine the breadth of microfluidic cell sorting technologies, while focusing on those that offer the greatest potential for translation into clinical and industrial practice and that offer multiple, useful functions. We organize these sorting technologies by the type of cell preparation required (i.e., fluorescent label-based sorting, bead-based sorting, and label-free sorting) as well as by the physical principles underlying each sorting mechanism.

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A monolithic glass chip for active single-cell sorting based on mechanical phenotyping

Christoph Faigle, Franziska Lautenschläger, Graeme Whyte, Philip Homewood, Estela Martín-Badosa and Jochen Guck

The mechanical properties of biological cells have long been considered as inherent markers of biological function and disease. However, the screening and active sorting of heterogeneous populations based on serial single-cell mechanical measurements has not been demonstrated. Here we present a novel monolithic glass chip for combined fluorescence detection and mechanical phenotyping using an optical stretcher. A new design and manufacturing process, involving the bonding of two asymmetrically etched glass plates, combines exact optical fiber alignment, low laser damage threshold and high imaging quality with the possibility of several microfluidic inlet and outlet channels. We show the utility of such a custom-built optical stretcher glass chip by measuring and sorting single cells in a heterogeneous population based on their different mechanical properties and verify sorting accuracy by simultaneous fluorescence detection. This offers new possibilities of exact characterization and sorting of small populations based on rheological properties for biological and biomedical applications.

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