RESUMO
Tissues contain exquisite vascular microstructures, and patterns of chemical cues for directing cell migration, homing, and differentiation for organ development and function. 3D microfabrication by multi-photon photolithography is a flexible, high-resolution tool for generating 3D bioscaffolds. However, the combined fabrication of scaffold microstructure simultaneously with patterning of cues to create both geometrically and chemically defined microenvironments remains to be demonstrated. This study presents a high-speed method for micron-resolution fabrication of scaffold microstructure and patterning of protein cues simultaneously using native scaffold materials. By the simultaneous microfabrication of arbitrary microvasculature geometries, and patterning selected regions of the microvasculature with the homing ligand P-selectin, this study demonstrates adhesion, rolling, and selective homing of cells in defined 3D regions. This novel ability to generate high-resolution geometries replete with patterned cues at high speed enables the construction of biomimetic microenvironments for complex 3D assays of cellular behavior.
Assuntos
Alicerces Teciduais/química , Materiais Biocompatíveis/química , Biomimética/métodos , Células Cultivadas , Células HL-60 , Humanos , Microtecnologia/métodos , Selectina-P/metabolismo , Fótons , Engenharia Tecidual/métodosRESUMO
Various microfluidic cell culture devices have been developed for in vitro cell studies because of their capabilities to reconstitute in vivo microenvironments. However, controlling flows in microfluidic devices is not straightforward due to the wide varieties of fluidic properties of biological samples. Currently, flow observations mainly depend on optical imaging and macro scale transducers, which usually require sophisticated instrumentation and are difficult to scale up. Without real time monitoring, the control of flows can only rely on theoretical calculations and numerical simulations. Consequently, these devices have difficulty in being broadly exploited in biological research. This paper reports a microfluidic device with embedded pressure sensors constructed using electrofluidic circuits, which are electrical circuits built by fluidic channels filled with ionic liquid. A microfluidic device culturing endothelial cells under various shear stress and hydrostatic pressure combinations is developed to demonstrate this concept. The device combines the concepts of electrofluidic circuits for pressure sensing, and an equivalent circuit model to design the cell culture channels. In the experiments, human umbilical vein endothelial cells (HUVECs) are cultured in the device with a continuous medium perfusion, which provides the combinatory mechanical stimulations, while the hydrostatic pressures are monitored in real time to ensure the desired culture conditions. The experimental results demonstrate the importance of real time pressure monitoring, and how both mechanical stimulations affect the HUVEC culture. This developed microfluidic device is simple, robust, and can be easily scaled up for high-throughput experiments. Furthermore, the device provides a practical platform for an in vitro cell culture under well-controlled and dynamic microenvironments.
Assuntos
Células Endoteliais da Veia Umbilical Humana , Técnicas Analíticas Microfluídicas , Modelos Biológicos , Estresse Fisiológico , Células Cultivadas , Eletrodos , Células Endoteliais da Veia Umbilical Humana/citologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Pressão Hidrostática , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos , Pressão , Resistência ao CisalhamentoRESUMO
Microfluidic technology plays an essential role in various lab on a chip devices due to its desired advantages. An automated microfluidic system integrated with actuators and sensors can further achieve better controllability. A number of microfluidic actuation schemes have been well developed. In contrast, most of the existing sensing methods still heavily rely on optical observations and external transducers, which have drawbacks including: costly instrumentation, professional operation, tedious interfacing, and difficulties of scaling up and further signal processing. This paper reports the concept of electrofluidic circuits - electrical circuits which are constructed using ionic liquid (IL)-filled fluidic channels. The developed electrofluidic circuits can be fabricated using a well-developed multi-layer soft lithography (MSL) process with polydimethylsiloxane (PDMS) microfluidic channels. Electrofluidic circuits allow seamless integration of pressure sensors with analog and digital operation functions into microfluidic systems and provide electrical readouts for further signal processing. In the experiments, the analog operation device is constructed based on electrofluidic Wheatstone bridge circuits with electrical outputs of the addition and subtraction results of the applied pressures. The digital operation (AND, OR, and XOR) devices are constructed using the electrofluidic pressure controlled switches, and output electrical signals of digital operations of the applied pressures. The experimental results demonstrate the designed functions for analog and digital operations of applied pressures are successfully achieved using the developed electrofluidic circuits, making them promising to develop integrated microfluidic systems with capabilities of precise pressure monitoring and further feedback control for advanced lab on a chip applications.
RESUMO
We demonstrate a novel, vertical temperature-mapping incubator utilizing eight layers of thermoelectric (TE) modules mounted around a test tube. The temperature at each layer of the TE module is individually controlled to simulate the vertical temperature profile of geo-temperature variations with depth. Owing to the constraint of non-intrusion to the filled geo-samples, the temperature on the tube wall is adopted for measurement feedback. The design considerations for the incubator include spatial arrangement of the energy transfer mechanism, heating capacity of the TE modules, minimum required sample amount for follow-up instrumental or chemical analysis, and the constraint of non-intrusion to the geo-samples during incubation. The performance of the incubator is experimentally evaluated with two tube conditions and under four preset temperature profiles. Test tubes are either empty or filled with quartz sand, which has comparable thermal properties to the materials in the geo-environment. The applied temperature profiles include uniform, constant temperature gradient, monotonic-increasing parabolic, and parabolic. The temperature on the tube wall can be controlled between 20 °C and 90 °C with an averaged root mean squared error of 1 °C.