Currently, research on the construction of vascular-like tubular structures is a hot area of tissue engineering, since it has potential applications in the building of artificial blood vessels. In this paper, we report a fluidic self-assembly method using cell embedded microstructures to construct vascular-like microtubes. A novel 4-layer microfluidic device was fabricated using polydimethylsiloxane (PDMS), which contains fabrication, self-assembly and extraction areas inside one channel. Cell embedded microstructures were directly fabricated using poly(ethylene glycol) diacrylate (PEGDA) in the fabrication area, namely on-chip fabrication. Self-assembly of the fabricated microstructures was performed in the assembly area which has a micro well. Assembled tubular structures (microtubes) were extracted outside the channel into culture dishes using a normally closed (NC) micro valve in the extraction area. The self-assembly mechanism was experimentally demonstrated. The performance of the NC micro valve and embedded cell concentration were both evaluated. Fibroblast (NIH/3T3) embedded vascular-like microtubes were constructed inside this reusable microfluidic device.

Fabricated vessel-mimetic microtubes are essential for delivering sufficient nutrient to engineered composite tissues. In this paper, vascular-like microtubes are engineered by automated assembly of donut-shaped micromodules that embed fibroblast cells. A microrobotic system is set up with dual manipulators of 30-nm positioning resolution under an optical microscope. The system assembles the micromodules by repeated single-step pick-up motions. This process is specifically designed to avoid human interference and ensure high reproducibility for automation. We optimized the single-step motion by calibrating the key parameters (the micromodule dimensions) in a force analysis. The optimal motion achieved a 98% pick-up success rate. The automated repetitive single-step assembly is achieved by an algorithm that acquires the 3-D location and tracks the micromanipulator without being affected by low contrast. The accuracy of the acquired 3-D location was experimentally determined as approximately 1 pixel (2 μm under 4× magnification), and the tracking under different observation conditions is proved effective. Finally, we automatically assembled microtubes at 6 micromodules/min, sufficiently fast for fabricating macroscopic vessel-mimetic substitutes in biological applications.

In this study, we introduce novel method of flow cytometry for cell detection based on impedance measurements. The state of the art method for impedance flow cytometry detection utilizes an embedded electrode in the microfluidic to perform measurement of electrical impedance of the presence of cells at the sensing area. Nonetheless, this method requires an expensive and complicated electrode fabrication process. Furthermore, reuse of the fabricated electrode also requires an intensive and tedious cleaning process. Due to that, we present a microfluidic device with integrated microneedles. The two microneedles are placed at the half height of the microchannel for cell detection and electrical measurement. A commercially-available Tungsten needle was utilized for the microneedles. The microneedles are easily removed from the disposable PDMS (Polydimethylsiloxane) microchannel and can be reused with a simple cleaning process, such as washing by ultrasonic cleaning. Although this device was low cost, it preserves the core functionality of the sensor, which is capable of detecting passing cells at the sensing area. Therefore, this device is suitable for low-cost medical and food safety screening and testing process in developing countries.