Team:BIT/HW1
Results
Ⅰ. Biological part:
Ⅱ. Fluids part:
Ⅲ. Optical part:
In our project, the chip is the carrier of biological reactions, its most basic function is to provide a growth environment for engineering bacteria. In order to be used smoothly as a biological carrier in our project, the chip needs to meet the following conditions:
①:The freeze-drying and recovery of engineering bacteria can be finished in the chip.And the engineering bacteria needs to keep a relatively high activity after recovery process.
Results:
①:The chip can ensure the normal growth of bacteria in a short time, but its long-term cultivating performance is not so good.According to our analysis,it may be due to the hypoxia environment in the chip.
②:The hypoxic environment does have a great impact on the growth of bacteria.
③:Freeze-drying and recovery process of engineering bacteria can be completed.
④:The bacteria remains relatively high activity after recovery.
In summary, as biological reaction carrier, the chip can be qualified in the short term bacterial culture,so the chip can meet our basic requirements.But the performance of long-term cultivating is poor.According to our analysis,it may be due to the internal hypoxic environment of the chip.Experiments have shown as proof that the hypoxic environment does have a great impact on the growth of bacteria.
As a microfluidic chip, the assessment of the flow of liquid is a necessary part of characterizing the performance of the chip. For the purposes of this project, the risks that may present during the infusion process include:
①: Blocking may occur in injection process.As a consequence,the injection would be not smooth,which would even lead to chip damage.
②: Bubbles are possibly be produced during injection,which would lead to dead volume increasing and biochemical reaction as well as optical detection process would be impacted consequently.
③: The fast flow rate may make the upstream beads to be washed to the downstream, so that the aptamer’s complementary strand would cleavage and release lysine with absence of the AFP,which would increase the risk of false positive diagnosis.
In contrast, in this regard we need to meet the requirements as follows:
①: Ability to perform a smooth injection without blocking and bubbles in constant injection rate condition.
②: Magnetic beads would not be washed to the downstream.
Results:
At 37 μl / min injection conditions, the chip does not produce bubbles and clogging, but there will be a small amount of beads being washed into the downstream chamber. How to eliminate this part of error led by these magnetic beads of is still need to considerate as the improvement of the project.
On the one hand, our chip is the carrier of biological reactions, on the other hand, the chip is also our optical detection platform.The object we are going to detect is GFP and RFP.
The detection principle is using a certain wavelength of light through the chip to stimulate the GFP and RFP in the chamber and detecting the light intensity emitted by GFP and RFP.In this way,we can characterize the amount of fluorescent protein produced by engineering bacteria.Thus the relationship between the reaction of engineering bacteria and the light intensity was established.
So the upper part of the chip will not only through the instrument from the excitation light, but also through the emission light from the engineering bacteria. If the chip has a poor transmittance, both the excitation light and light will be seriously attenuated.Thus it will affect the detection accuracy.
Results:
The result indicates that light transmittance of our chip is ideal enough to ignore the error caused by light attenuation.
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