Team:BostonU HW/Improve

Improving Upon the 2016 BostonU_HW Project: Neptune

During the course of our 2017 iGEM Project, the BostonU_HW team worked to improve upon the 2016 BostonU_HW Project: Neptune. Neptune allowed uses to specify, design, and control a microfluidic chip. For more information regarding specific elements within Neptune, please refer to their wiki. Our team worked to improve the specify and design portion of the Neptune software workflow. This Neptune can be broken down into five basic blocks:

The portion of Neptune’s workflow we contributed to was Fluigi. Fluigi is a place-and-route software that receives a detailed microfluidic description in the form of MINT code, and outputs a fully routed chip design with accompanying SVG and JSON files. During the process of our iGEM project, 17 distinct microfluidic chip designs were coded in MINT and routed using Fluigi. These chips ranged from designs replicated from various journals to original designs for the MARS Repository. While designing and routing these chips we were able to identify issues, limitations, and areas for improvement in the Fluigi and Neptune software system. These issues were identified as elements of Fluigi to fix or improve upon. During the course of our 2017 iGEM project, we altered design rules that would be necessary to integrate into the layout algorithms, some of them in particular are:

1. Ports are no longer automatically pushed to the edges of a design. When manufacturing a chip using photolithography ports must be at the edges of the chip. However, when using the Makerfluidics workflow this is no longer a requirement; ports can be located anywhere on a chip, within reason. In fact, many designs benefit from or rely on ports being placed in nontraditional locations. Therefore, the pre-existing design rule that ports must be pushed to the edges of a chip needed to be updated. Instead of automatically pushing all ports to the edges of a design, Fluigi can now infer whether or not doing so would be beneficial/required. Thus, the design rule was updated to be more relaxed and accurate for the Makerfluidics workflow.
2. Primitive locations can be set in the X and/or Y direction. When designing a chip using Fluigi’s place and route system, primitives are defined by the user but the location and connections of these elements of these primitives are determined by Fluigi’s place and route system. However, for some designs the location of primitives must be in specific locations. Fluigi contains a function that allows primitives to be locked in place, the set function, however this function requires both an X and Y coordinate. For some designs, only one of these coordinates is known or required. To accommodate this, Fluigi’s pre-existing “set” design rules was updated to allow the user to set a primitive’s X coordinate, Y coordinate, or both. This updated design rule made the “set” function more practical to use.
3. Subtle routing changes were added to make designs more practical and functional. Fluigi’s place and route system is effective at connecting primitives with channels, however, it did not always make these connections in the most practical or functional ways. For example, random bends or “kinks” would sometimes be added to channels during the routing process. While a chip with random bends is still a valid design, it may not necessarily be practical as these bends may affect fluid flow. Fluigi’s design rules regarding bend tolerance have been progressively increase over time, thus limiting the amount of unnecessary bends that can be added to a design.

The impact of these improved design rules can be seen in the two Fluigi-routed chips below. On the left is an output design for eColiBasic5. This chip was designed and routed prior to many of these improvements being implemented. While this chip represents a valid design, it is not optimized or practical due to bends and port location issues. On the right is an output design for eColiBasic6 following the implementation of the previously identified improvements. The MINT code for these two microfluidic chips is nearly identical, but the outputs are entirely different. eColiBasic6 is a cleaner, more practical and functional design that can be manufactured and tested.

Video Tutorials | Sarah and Dinithi

In order to make the microfluidic chips we designed accessible to synthetic biologists we developed four fully-narrated tutorial videos. These videos go step by step through the process of manufacturing, assembling, and testing a microfluidic device. The four videos created teach uses how to mill a microfluidic chip, how to make PDMS, how to assemble a microfluidic chip, and how to clean a microfluidic chip. Each of these videos is accompanied by a detailed written protocol as well.