Team:BIT/Design/HW
We chose the microfluidic chip because it has three significant advantages in the application:Microfluidic chip
1.The tiny volume of the microfluidic chip allows the liquid reaction system to be miniaturized and integrated. It can use a small amount of liquid and space to achieve complex biochemical reactions;
2.Reactions on the microfluidic chip can be quantitative analysis and detection;
3. Microfluidic chip can be combined with external devices to make biochemical reactions become automated and intelligent.
Based on the characteristics of microfluidic chip, more and more people use microfluidic chip as the platform of biochemical detection and POCT [1]. For example, there has been a microfluidic platform which can detect C-reactive protein in 2008,and the limit of detection is 2.6 ng/ml.Besides,it can be produced in large quantities[2].Similarly, in the case of hepatotoxicity assessment, a POCT method based on microfluidic systems overcomes the shortcomings of long processing times and high levels of personnel in traditional methods[3].It is worth mentioning that there already have mature POCT equipments based on microfluidic platform in the market.It can be seen that the microfluidic platform not only has irreplaceable performance advantages, but also has marketization potential.
Ⅰ.Design
An important feature of microfluidic chips is that they are capable of artificially designing liquid lines and reaction chambers. Our project involves three reaction processes: the separation of the aptamer and the complementary strand on the magnetic beads, the separation of the complementary strand from the small molecule, and the biochemical reaction of the small molecule and the engineered bacteria. According to the needs of the project, we designed the two reaction chambers as shown in Figure 1.1, and used the peristaltic pump, the magnet plate and the heating plate as supporting auxiliary equipment.
As can be seen from Figure 1.2, our chip consists of two chambers:
Chamber 1 is oval(long axis length:18mm, short axis length:6mm). In our design, the function of the chamber is to hold our magnetic beads (fixed with magnets) with the aptamer and provide a place for the sample to bind to the aptamer so that the the complementary strands with lysine can get detached . Our intention to design the chamber as an oval is that the elongated structure can reduce the bubble generated by the difference in flow rate between the cavity wall and the middle stream.
Chamber 2 is round( diameter:12mm) Within this chamber ,there are gel pillars(diameter: 0.5mm). The contents of the chamber are engineering bacteria frozen into dry powdery and trypsin. After the reaction in the chamber is carried out for a certain period of time, the mixed reaction of the culture medium and the reaction liquid flows from the upstream into the chamber under the negative pressure generated by the peristaltic pump (see the equipment section), the following reaction occurs:
①: Freeze-dried engineering bacteria’s recovery with the help of culture medium and constant temperature provided by the heating plate
②: Trypsin catalyzes cracking of DNA chain,lysine on the complementary strand falls into free small molecule
③: Lysine goes into the engineering bacteria, biochemical reaction within the engineering bacteria began.
We can use the design of hardware to detect the reaction generated fluorescence generated in reaction③, and we can draw a conclusion through analyzing and processing.
Ⅱ.Materials
The material we used to fabricate the chip is NOA 81.In normal conditions it’s in liquid state.It will solidify with exposure of 320nm to 380nm wavelength UV light.
We made the chip using the material NOA 81. The material is normally liquid, and when subjected to UV light from the 320nm to 380nm band, the material is quickly cured to form a solid with good hardness and toughness. [4] According to this characteristic, we can use soft lithography method to achieve the production of the chip . The reason why we used NOA81 as the material of our chip is not just it’s UV curing characteristics, the light transmittance of this material is also taken into consideration. The transmittance provided by the manufacturer is shown in Figure 1.3. At the same time, we also designed a verification experiment for the light transmission of the chip(can be found in demonstrate section )
Ⅲ.Working Process
Briefly,our chip went through following process between fabrication and finishing detection(shown in Figure 1.4):
①:Injecting magnetic beads to Chamber 1,then freeze-drying the liquid.
②:Injecting engineering bacteria to Chamber 2 ;then freeze-drying the liquid.Then the chip is ready to use.
③:When needing detection,taking the chip,connecting the peristaltic pump and injecting a volume of sample and culture medium mixture.
④:After the reaction is complete, the peristaltic pump pumps the reaction solution into the chamber with negative pressure. The biochemical reaction takes place and enters the subsequent detection process.
Reference
[1]:Mazher-Iqbal Mohammed and Marc P. Y. Desmulliez,Lab Chip, 2011,11, 569-595
[2]:C. Jonsson, M. Aronsson, G. Rundstrom, C. Pettersson, I. Medel-
Hartvig, J. Bakker, E. Martinsson, B. Liedberg, B. MacCraith,
O. Ohman and J. Melin, Lab Chip, 2008, 8, 1191–1197
[3]:Geok Soon Lim,Joseph S. Chang, Zhang Lei,Ruige Wu, Zhiping Wang, Kemi Cui and Stephen Wong,Lab Chip, 2015,15, 4032-4043
[4]:http://www.norlandprod.com/adhesives/noa%2081.html
