Zida Li

Emerging microengineered tools for functional analysis and phenotyping of blood cells

The available techniques for assessing blood cell functions are limited considering the various types of blood cell and their diverse functions. In the past decade, rapid advances in microengineering have enabled an array of blood cell functional measurements that are difficult or impossible to achieve using conventional bulk platforms. Such miniaturized blood cell assay platforms also provide the attractive capabilities of reducing chemical consumption, cost, and assay time, as well as exciting opportunities for device integration, automation, and assay standardization. This review summarizes these contemporary microengineered tools and discusses their promising potential for constructing accurate in vitro models and rapid clinical diagnosis using minimal amounts of whole-blood samples.

Musical interfaces: visualization and reconstruction of music with a microfluidic two-phase flow.

Detection of sound wave in fluids can hardly be realized because of the lack of approaches to visualize the very minute sound-induced fluid motion. In this paper, we demonstrate the first direct visualization of music in the form of ripples at a microfluidic aqueous-aqueous interface with an ultra-low interfacial tension. The interfaces respond to sound of different frequency and amplitude robustly with sufficiently precise time resolution for the recording of musical notes and even subsequent reconstruction with high fidelity. Our work shows the possibility of sensing and transmitting vibrations as tiny as those induced by sound. This robust control of the interfacial dynamics enables a platform for investigating the mechanical properties of microstructures and for studying frequency-dependent phenomena, for example, in biological systems.

Acoustic tweezing cytometry enhances osteogenesis of human mesenchymal stem cells through cytoskeletal contractility and YAP activation

Human mesenchymal stem cells (hMSCs) have great potential for cell-based therapies for treating degenerative bone diseases. It is known that mechanical cues in the cell microenvironment play an important role in regulating osteogenic (bone) differentiation of hMSCs. However, mechanoregulation of lineage commitment of hMSCs in conventional two-dimensional (2D) monocultures or bioengineered three-dimensional (3D) tissue constructs remains suboptimal due to complex biomaterial design criteria for hMSC culture. In this study, we demonstrate the feasibility of a novel cell mechanics and mechanobiology tool, acoustic tweezing cytometry (ATC), for mechanical stimulation of hMSCs. ATC utilizes ultrasound (US) pulses to actuate functionalized lipid microbubbles (MBs) which are covalently attached to hMSCs via integrin binding to exert forces to the cells. ATC stimulation increases cytoskeletal contractility of hMSCs regardless of the cell area. Furthermore, ATC application rescues osteogenic differentiation of hMSCs in culture conditions that are intrinsically repressive for hMSC osteogenesis (e.g., soft cell culture surfaces). ATC application activates transcriptional regulator YAP to enhance hMSC osteogenesis. Our data further show that F-actin, myosin II, and RhoA/ROCK signaling functions upstream of YAP activity in mediating ATC-stimulated hMSC osteogenesis. With the capability of applying controlled dynamic mechanical stimuli to cells, ATC provides a powerful tool for mechanoregulation of stem cell behaviors in tissue engineering and regenerative medicine applications.

A Miniaturized Hemoretractometer for Blood Clot Retraction Testing

Blood coagulation is a critical hemostatic process that must be properly regulated to maintain a delicate balance between bleeding and clotting. Disorders of blood coagulation can expose patients to the risk of either bleeding disorders or thrombotic diseases. Coagulation diagnostics using whole blood is very promising for assessing the complexity of the coagulation system and for global measurements of hemostasis. Despite the clinic values that existing whole blood coagulation tests have demonstrated, these systems have significant limitations that diminish their potential for point-of-care applications. Here, recent advancements in device miniaturization using functional soft materials are leveraged to develop a miniaturized clot retraction force assay device termed mHemoRetractoMeter (mHRM). The mHRM is capable of precise measurements of dynamic clot retraction forces in real time using minute amounts of whole blood. To further demonstrate the clinical utility of the mHRM, systematic studies are conducted using the mHRM to examine the effects of assay temperature, treatments of clotting agents, and pro- and anti-coagulant drugs on clot retraction force developments of whole blood samples. The mHRMs low fabrication cost, small size, and consumption of only minute amounts of blood samples make the technology promising as a point-of-care tool for future coagulation monitoring.

Tracking the tumor invasion front using long-term fluidic tumoroid culture

The analysis of invading leader cells at the tumor invasion front is of significant interest as these cells may possess a coordinated functional and molecular phenotype which can be targeted for therapy. However, such analyses are currently limited by available technologies. Here, we report a fluidic device for long-term three-dimensional tumoroid culture which recapitulated the tumor invasion front, allowing for both quantification of invasive potential and molecular characterization of invasive leader cells. Preliminary analysis of the invasion front indicated an association with cell proliferation and higher expression of growth differentiation factor 15 (GDF15). This device makes real-time tracking of invading leader cell phenotypes possible and has potential for use with patient material for clinical risk stratification and personalized medicine.

Carbon Nanotube Strain Sensor Based Hemoretractometer for Blood Coagulation Testing

Coagulation monitoring is essential for perioperative care and thrombosis treatment. However, existing assays for coagulation monitoring have limitations such as a large footprint and complex setup. In this work, we developed a miniaturized device for point-of-care blood coagulation testing by measuring dynamic clot retraction force development during blood clotting. In this device, a blood drop was localized between a protrusion and a flexible force-sensing beam to measure clot retraction force. The beam was featured with micropillar arrays to assist the deposition of carbon nanotube films, which served as a strain sensor to achieve label-free electrical readout of clot retraction force in real time. We characterized mechanical and electrical properties of the force-sensing beam and optimized its design. We further demonstrated that this blood coagulation monitoring device could obtain results that were consistent with those using an imaging method and that the device was capable of differentiating blood samples with different coagulation profiles. Owing to its low fabrication cost, small size, and low consumption of blood samples, the blood coagulation testing device using carbon nanotube strain sensors holds great potential as a point-of-care tool for future coagulation monitoring.

Capillary assisted deposition of carbon nanotube film for strain sensing

Advances in stretchable electronics offer the possibility of developing skin-like motion sensors. Carbon nanotubes (CNTs), owing to their superior electrical properties, have great potential for applications in such sensors. In this paper, we report a method for deposition and patterning of CNTs on soft, elastic polydimethylsiloxane (PDMS) substrates using capillary action. Micropillar arrays were generated on PDMS surfaces before treatment with plasma to render them hydrophilic. Capillary force enabled by the micropillar array spreads CNT solution evenly on PDMS surfaces. Solvent evaporation leaves a uniform deposition and patterning of CNTs on PDMS surfaces. We studied the effect of the CNT concentration and micropillar gap size on CNT coating uniformity, film conductivity, and piezoresistivity. Leveraging the piezoresistivity of deposited CNT films, we further designed and characterized a device for the contraction force measurement. Our capillary assisted deposition method of CNT films showed great application potential in fabrication of flexible CNT thin films for strain sensing.

Clot Retraction: A Miniaturized Hemoretractometer for Blood Clot Retraction Testing (Small 29/2016).

Whole blood coagulation testing provides valuable diagnostic information on diseases such as bleeding disorders, heart attack, deep venous thrombosis, etc. On page 3926, J. Fu and co-workers develop a miniaturized hemoretractometer to measure clot contraction upon blood coagulation with good reproducibility and robustness. This device design shows great application potential in point-of-care testing. Photo credit: David Peyer from University of Michigan.