Liyu Liu

Deterministic separation of cancer cells from blood at 10 mL/min

Circulating tumor cells (CTCs) and circulating clusters of cancer and stromal cells have been identified in the blood of patients with malignant cancer and can be used as a diagnostic for disease severity, assess the efficacy of different treatment strategies and possibly determine the eventual location of metastatic invasions for possible treatment. There is thus a critical need to isolate, propagate and characterize viable CTCs and clusters of cancer cells with their associated stroma cells. Here, we present a microfluidic device for mL/min flow rate, continuous-flow capture of viable CTCs from blood using deterministic lateral displacement (DLD) arrays. We show here that a DLD array device can isolate CTCs from blood with capture efficiency greater than 85% CTCs at volumetric flow rates of up to 10 mL/min with no effect on cell viability.

Design and integration of an all-in-one biomicrofluidic chip

We demonstrate a highly integrated microfluidic chip with the function of DNA amplification. The integrated chip combines giant electrorheological-fluid actuated micromixer and micropump with a microheater array, all formed using soft lithography. Internal functional components are based on polydimethylsiloxane (PDMS) and silvercarbon black-PDMS composites. The system has the advantages of small size with a high degree of integration, high polymerase chain reaction efficiency, digital control and simple fabrication at low cost. This integration approach shows promise for a broad range of applications in chemical synthesis and biological sensinganalysis, as different components can be combined to target desired functionalities, with flexible designs of different microchips easily realizable through soft lithography.

Active microfluidic mixer chip

We report the design and fabrication of a chaotic mixer based on the electrorheological (ER) fluid-controlled valves. The flow in the main channel is perturbed by liquid flow in orthogonal side channels, driven by hydrodynamic pulsating pumps. Each pulsating pump consists of a chamber with diaphragm plus two out-of-phase ER valves operating in a push-pull mode. All the valves, pumps, and mixing channels are integrated in one polydimethylsioxane chip. Mixing characteristics in the main channel are controlled by the strength and frequency of external electric fields applied on the ER fluid.

Paperlike thermochromic display

The authors report the design and implementation of a paperlike, thermally activated display fabricated from thermochromic composite and embedded conductive wiring patterns, shaped from mixture of metallic nanoparticles in polydimethylsioxane using soft lithography. The display exhibits good image quality and ease of control. Use of electric heating pulses is shown to reduce energy consumption while improving image quality control. The display has excellent mechanical bending flexibility.

Microheaters fabricated from a conducting composite

The authors report the fabrication of microheaters based on a conducting composite of silver microparticles embedded in polydimethylsioxane (PDMS). Experimental results show that the microheaters exhibit good performance in temperature rise and decay characteristics, with localized heating at targeted spatial domains. A unique feature of the microheater here is its excellent integration capability with biocompatible PDMS and other polymer materials, with potential microchip applications for bioprocessing and chemical reactions.

Electrorheological Fluid-actuated Microfluidic Pump

The authors report the design and implementation of an electrorheological (ER) fluid-actuated microfluidic pump, with programmable digital control. Our microfluidic pump has a multilayered structure fabricated on polydimethylsiloxane by soft-lithographic technique. The ER microfluidic pump exhibits good performance at high pumping frequencies and uniform liquid flow characteristics. It can be easily integrated with other microfluidic components. The programmable control also gives the device flexibility in its operations.

Minimization of thermodynamic costs in cancer cell invasion

Metastasis, the truly lethal aspect of cancer, occurs when metastatic cancer cells in a tumor break through the basement membrane and penetrate the extracellular matrix. We show that MDA-MB-231 metastatic breast cancer cells cooperatively invade a 3D collagen matrix while following a glucose gradient. The invasion front of the cells is a dynamic one, with different cells assuming the lead on a time scale of 70 h. The front cell leadership is dynamic presumably because of metabolic costs associated with a long-range strain field that precedes the invading cell front, which we have imaged using confocal imaging and marker beads imbedded in the collagen matrix. We suggest this could be a quantitative assay for an invasive phenotype tracking a glucose gradient and show that the invading cells act in a cooperative manner by exchanging leaders in the invading front.

Oriented collagen fibers direct tumor cell intravasation

Significance Intravasation is an early stage of metastasis that involves metastatic cells moving from the tumor into the extracellular matrix (ECM), breakthrough of the basement membrane, and entry into blood vessels. We found that the oriented fibers greatly enhance and facilitate the metastatic cell intravasation process during metastasis. We suggest that a possible “tissue treatment” therapy could be considered, in which the ECM fiber structure orientation in the tumor microenvironment might be altered to minimize the intravasation rate of metastatic cells. In this work, we constructed a Collagen I–Matrigel composite extracellular matrix (ECM). The composite ECM was used to determine the influence of the local collagen fiber orientation on the collective intravasation ability of tumor cells. We found that the local fiber alignment enhanced cell–ECM interactions. Specifically, metastatic MDA-MB-231 breast cancer cells followed the local fiber alignment direction during the intravasation into rigid Matrigel (∼10 mg/mL protein concentration).

Probing the invasiveness of prostate cancer cells in a 3D microfabricated landscape

The metastatic invasion of cancer cells from primary tumors to distant ecological niches, rather than the primary tumors, is the cause of much cancer mortality [Zhang QB, et al. (2010) Int J Cancer 126:2534–2541; Chambers AF, Goss PE (2008) Breast Cancer Res 10:114]. Metastasis is a three-dimensional invasion process where cells spread from their site of origin and colonize distant microenvironmental niches. It is critical to be able to assess quantitatively the metastatic potential of cancer cells [Harma V, et al. (2010) PLoS ONE 5:e10431]. We have constructed a microfabricated chip with a three-dimensional topology consisting of lowlands and isolated square highlands (Tepuis), which stand hundreds of microns above the lowlands, in order to assess cancer cell metastatic potential as they invade the highlands. As a test case, the invasive ascents of the Tepui by highly metastatic PC-3 and noninvasive LNCaP prostate cancer cells were used. The vertical ascent by prostate cancer cells from the lowlands to the tops of the Tepui was imaged using confocal microscopy and used as a measure of the relative invasiveness. The less-metastatic cells (LNCaP) never populated all available tops, leaving about 15% of them unoccupied, whereas the more metastatic PC-3 cells occupied all available Tepuis. We argue that this distinct difference in invasiveness is due to contact inhibition.

A microfluidic device for continuous cancer cell culture and passage with hydrodynamic forces

We demonstrate a novel and robust microfluidic chip with combined functions of continuous culture and output of PC-3 prostate cancer cells. With digital controls, polydimethylsiloxane (PDMS) flexible diaphragms are able to apply hydrodynamic shear forces on cultures, detaching a fraction of attached cancer cells from the surface for output while leaving others for reuse in subsequent cultures. The fractions of detached cells and remaining cells can be precisely controlled. The system has not only the advantages of small size, high cell culture efficiency, and digital control, but also of simple fabrication at low cost, easy operation and robust performance. The chip performs 9 passages during 30 days of continuous culture and shows promise as a durable design suitable for long-term cell output.