Three-dimensional lab-on-a-foil device for dielectrophoretic separation of cancer cells.

A simple, low-cost, three-dimensional (3D) lab-on-a-foil microfluidic device for dielectrophoretic separation of circulating tumor cells (CTCs) is designed and constructed. Disposable thin films are cut by xurography and microelectrode array are made with rapid inkjet printing. The multilayer device design allows the studying of spatial movements of CTCs and red blood cells (RBCs) under dielectrophoresis (DEP). A numerical simulation was performed to find the optimum driving frequency of RBCs and the crossover frequency for CTCs. At the optimum frequency, RBCs were lifted 120 µm in z-axis direction by DEP force, and CTCs were not affected due to negligible DEP force. By utilizing the displacement difference, the separation of CTCs (modeled with A549 lung carcinoma cells) from RBCs in z-axis direction was achieved. With the nonuniform electric field at optimized driving frequency, the RBCs were trapped in the cavities above the microchannel, whereas the A549 cells were separated with a high capture rate of 86.3% ± 0.2%. The device opens not only the possibility for 3D high-throughput cell separation but also for future developments in 3D cell manipulation through rapid and low-cost fabrication.

Hierarchical data visualization of experimental erythrocyte aggregation employing cross correlation and optical flow applications.

Appraisal of microvascular erythrocyte velocity as well as aggregation are critical features of hemorheological assessment. Examination of erythrocyte velocity-aggregate characteristics is critical in assessing disorders associated with coagulopathy. Microvascular erythrocyte velocity can be assessed using various methodologic approaches; however, the shared assessment of erythrocyte velocity and aggregation has not been well described. The purpose of this study therefore is to examine three independent erythrocyte assessment strategies with and without experimentally induced aggregation in order to elucidate appropriate analytic strategy for combined velocity/aggregation assessment applicable to in-vivo capillaroscopy. We employed a hierarchical microfluidic model combined with Bland-Altman analysis to examine agreement between three methodologies to assess erythrocyte velocity appropriate for interpretation of cinematography of in-vivo microvascular hemorheology. We utilized optical and manual techniques as well as a technique which we term transversal temporal cross-correlation (TTC) to observe and measure both erythrocyte velocity and aggregation. In general, optical, manual and TTC agree in estimation of velocity at relatively low flow rate, however with an increase in infusion rate the optical flow method yielded the velocity estimates that were lower than the TTC and manual velocity estimates. We suggest that this difference was due to the fact that slower moving particles close to the channel wall were better illuminated than faster particles deeper in the channel which affected the optical flow analysis. Combined velocity/aggregation appraisal using TTC provides an efficient approach for estimating erythrocyte aggregation appropriate for in-vivo applications. We demonstrated that the optical flow and TTC analyses can be used to estimate erythrocyte velocity and aggregation both in ex-vivo microfluidics laboratory experiments as well as in-vivo recordings. The simplicity of TTC method may be advantageous for developing velocity estimate methods to be used in the clinic. The trade-off is that TTC estimation cannot capture features of the flow based on optical flow analysis of individually tracked particles.

Acoustic bubble for spheroid trapping, rotation, and culture: a tumor-on-a-chip platform (ABSTRACT platform).

Cancer is the leading cause of death globally, with 90% of deaths being caused by cancer metastasis. Circulating tumor cells (CTCs) play an important role in early diagnosis of cancer metastasis and in monitoring of therapeutic response. Therefore, reliable methods to isolate, collect and culture CTCs are required to obtain information on metastasis status and therapeutic treatment. In this work, we present a CTC-processing system: acoustic bubble for spheroid trapping, rotation, and culture: a tumor-on-a-chip platform (ABSTRACT). The platform consists of a main channel, several parallel sub-microchannels with microcavities and culture chambers. The microcavity is designed to trap a bubble with desired shape at the entrance of the sub-microchannel. Under the acoustic actuation, the trapped bubble oscillates and creates a secondary radiation force to trap and rotate CTCs at a desired location. By controlling the acoustic bubble, CTCs can be continuously trapped from the blood flow, rotated to form a spheroid, and released to the microchamber for culture. We systematically investigated the effects of device geometry, flow parameters, and input voltage on trapping of CTCs to optimize the performance. Additionally, the successful on-chip spheroid culture demonstrates the biocompatibility and the simplicity of this platform. Besides simplifying conventional complex CTC processing procedures, this ABSTRACT platform also shows great potential for downstream analysis of tumor cells, such as monitoring the progression of metastasis and personalized drug testing.

Study of ultrasound thrombolysis using acoustic bubbles in a microfluidic device.

Thrombosis is a common medical entity associated with many forms of cardiovascular disease including myocardial infarction and stroke. Recently, ultrasound thrombolysis has emerged as a promising technique for thrombosis treatment by delivering acoustic waves onto blood clots. In this study, an ultrasound thrombolysis method is presented using an acoustic bubble-based microfluidic device. With acoustic actuation, microstreaming flow is created in the microchannel by oscillating bubbles, breaking up the blood clots in blood samples in a few milliseconds. In a low-frequency field, the effects of bubble size on microstreaming patterns and thrombolysis have been experimentally studied. Using image processing techniques, we have quantitatively investigated the relationship between the input signal and the thrombolysis performance. Additionally, the viability test proved that there are no significant detrimental effects on the blood cells after acoustic actuation. This acoustic bubble-based microfluidic device is demonstrated to be a promising platform for quantitative analysis of ultrasound thrombolysis. It opens up possibilities for future development of ultrasound thrombolysis devices for the diagnosis and treatment of heart diseases.

Trapping and control of bubbles in various microfluidic applications.

As a simple, clean and effective tool, micro bubbles have enabled advances in various lab on a chip (LOC) applications recently. In bubble-based microfluidic applications, techniques for capturing and controlling the bubbles play an important role. Here we review active and passive techniques for bubble trapping and control in microfluidic applications. The active techniques are categorized based on various types of external forces from optical, electric, acoustic, mechanical and thermal fields. The passive approaches depend on surface tension, focusing on optimization of microgeometry and modification of surface properties. We discuss control techniques of size, location and stability of microbubbles and show how these bubbles are employed in various applications. To finalize, by highlighting the advantages of these approaches along with the current challenges, we discuss the future prospects of bubble trapping and control in microfluidic applications.

Acoustic Microfluidic Separation Techniques and Bioapplications: A Review.

Microfluidic separation technology has garnered significant attention over the past decade where particles are being separated at a micro/nanoscale in a rapid, low-cost, and simple manner. Amongst a myriad of separation technologies that have emerged thus far, acoustic microfluidic separation techniques are extremely apt to applications involving biological samples attributed to various advantages, including high controllability, biocompatibility, and non-invasive, label-free features. With that being said, downsides such as low throughput and dependence on external equipment still impede successful commercialization from laboratory-based prototypes. Here, we present a comprehensive review of recent advances in acoustic microfluidic separation techniques, along with exemplary applications. Specifically, an inclusive overview of fundamental theory and background is presented, then two sets of mechanisms underlying acoustic separation, bulk acoustic wave and surface acoustic wave, are introduced and discussed. Upon these summaries, we present a variety of applications based on acoustic separation. The primary focus is given to those associated with biological samples such as blood cells, cancer cells, proteins, bacteria, viruses, and DNA/RNA. Finally, we highlight the benefits and challenges behind burgeoning developments in the field and discuss the future perspectives and an outlook towards robust, integrated, and commercialized devices based on acoustic microfluidic separation.

Acoustofluidic stick-and-play micropump built on foil for single-cell trapping.

The majority of microfluidic devices nowadays are built on rigid or bulky substrates such as glass slides and polydimethylsiloxane (PDMS) slabs, and heavily rely on external equipment such as syringe pumps. Although a variety of micropumps have been developed in the past, few of them are suitable for flexible microfluidics or lab-on-a-foil systems. In this paper, stick-and-play acoustic micropump is built on thin and flexible plastic film by printing microstructures termed defended oscillating membrane equipped structures (DOMES) using two-photon polymerization. Specifically, this new micropump induces rectified flow upon the actuation of acoustic waves, and the flow patterns agree with simulation results very well. More importantly, the developed micropump has the capabilities to generate adjustable flow rates as high as 420 nL min-1, and does not suffer from problems such as bubble instability, gas dissolution, and undesired bubble-trapping that commonly occur in other forms of acoustic micropumps. Since the micropump works in stick-and-play mode, it is reusable after cleaning thanks to the easy separation of covers and substrates. Lastly, the developed micropump is applied for creating a self-pumped single-cell trapping device. The excellent trapping capability of the integrated device proves its potential for long-term studies of biological behaviors of individual cells for biomedical applications.