Micromanipulation of Live Microdissected Tissues with a Low-Cost Integrated Robotic Platform.

The small amount of human tissue available for testing is a paramount challenge in cancer drug development, cancer disease models, and personalized oncology. Technologies that combine the microscale manipulation of tissues with fluid handling offer the exciting possibility of miniaturizing and automating drug evaluation workflows. This approach minimizes animal testing and enables inexpensive, more efficient testing of samples with high clinical biomimicry using scarce materials. We have developed an inexpensive platform based on an off-the-shelf robot that can manipulate microdissected tissues (µDTs) into user-programmed positions without using intricate microfluidic designs nor any other accessories such as a microscope or a pneumatic controller. The robot integrates complex functions such as vision and fluid actuation by incorporating simple items including a USB camera and a rotary pump. Through the robot's camera, the platform software optically recognizes randomly-seeded µDTs on the surface of a petri dish and positions a mechanical arm above the µDTs. Then, a custom rotary pump actuated by one of the robot's motors generates enough microfluidic lift to hydrodynamically pick and place µDTs with a pipette at a safe distance from the substrate without requiring a proximity sensor. The platform's simple, integrated construction is cost-effective and compact, allowing placement inside a tissue culture hood for sterile workflows. The platform enables users to select µDTs based on their size, place them in user-programmed arrays, such as multi-well plates, and control various robot motion parameters. As a case application, we use the robotic system to conduct semi-automated drug testing of mouse and human µDTs in 384-well plates. Our user-friendly platform promises to democratize microscale tissue research to clinical and biological laboratories worldwide.

Tunable resins with PDMS-like elastic modulus for stereolithographic 3D-printing of multimaterial microfluidic actuators.

Stereolithographic 3D-printing (SLA) permits facile fabrication of high-precision microfluidic and lab-on-a-chip devices. SLA photopolymers often yield parts with low mechanical compliancy in sharp contrast to elastomers such as poly(dimethyl siloxane) (PDMS). On the other hand, SLA-printable elastomers with soft mechanical properties do not fulfill the distinct requirements for a highly manufacturable resin in microfluidics (e.g., high-resolution printability, transparency, low-viscosity). These limitations restrict our ability to print microfluidic actuators containing dynamic, movable elements. Here we introduce low-viscous photopolymers based on a tunable blend of the monomers poly(ethylene glycol) diacrylate (PEGDA, Mw ∼ 258) and the monoacrylate poly(ethylene glycol methyl ether) methacrylate (PEGMEMA, Mw ∼ 300). In these blends, which we term PEGDA-co-PEGMEMA, tuning the PEGMEMA content from 0% to 40% (v/v) alters the elastic modulus of the printed plastics by ∼400-fold, reaching that of PDMS. Through the addition of PEGMEMA, moreover, PEGDA-co-PEGMEMA retains desirable properties of highly manufacturable PEGDA such as low viscosity, solvent compatibility, cytocompatibility and low drug absorptivity. With PEGDA-co-PEGMEMA, we SLA-printed drastically enhanced fluidic actuators including microvalves, micropumps, and microregulators with a hybrid structure containing a flexible PEGDA-co-PEGMEMA membrane within a rigid PEGDA housing. These components were built using a custom "Print-Pause-Print" protocol, referred to as "3P-printing", that allows for fabricating high-resolution multimaterial parts with a desktop SLA printer without the need for post-assembly. SLA-printing of multimaterial microfluidic actuators addresses the unmet need of high-performance on-chip controls in 3D-printed microfluidic and lab-on-a-chip devices.

A 'print-pause-print' protocol for 3D printing microfluidics using multimaterial stereolithography.

Methods to make microfluidic chips using 3D printers have attracted much attention because these simple procedures allow rapid fabrication of ready-to-use products from digital 3D designs with minimal human intervention. Printing high-resolution chips that are simultaneously transparent, biocompatible and contain regions of dissimilar materials is an ongoing challenge. Transparency allows for the optical inspection of specimens containing cells and labeled biomolecules inside the chip. Being able to use different materials for different layers in the product increases the number of potential applications. In this 'print-pause-print' protocol, we describe detailed strategies for fabricating transparent biomicrofluidic devices and multimaterial chips using stereolithographic 3D printing. To print transparent biomicrofluidic chips, we developed a transparent resin based on poly(ethylene glycol) diacrylate (PEG-DA) (average molecular weight: 250 g/mol, PEG-DA-250) and a smooth chip surface technique achieved using glass. Cells can be successfully cultured and visualized on PEG-DA-250 prints and inside PEG-DA-250 microchannels. The multimaterial potential of the technique is exemplified using a molecular diffusion device that comprises parts made of two different materials: the channel walls, which are water impermeable, and a porous barrier structure, which is permeable to small molecules that diffuse through it. The two materials were prepared from two different molecular-weight PEG-DA-based printing resins. Alignment of the two dissimilar material structures is performed automatically by the printer during the printing process, which only requires a simple pause step to exchange the resins. The procedure takes less than 1 h and can facilitate chip-based applications including biomolecule analysis, cell biology, organ-on-a-chip and tissue engineering.