Team:Tsinghua/Measurement
Measurement
Overview
This year, NBA attempts to induce the dispersal of biofilm by our engineered NO-releasing bacteria. The equipment listed below were used to test the solidity of our system of.
Microplate Reader
Varioskan LUX
The microplate reader was used primarily for the aim to detect relative fluorescence unites (RFU) in several systems.
As we have constructed strains that can express EGFP when induced, it is promising to use the microplate reader to visualize the condition of our biofilm. RFU was calculated at the process of cultivation biofilms and after the biofilm was treated with SNP, the NO donor.
In detecting the working efficiency of our C4-HSL-EGFP report system and phmp-EGFP system, the microplate reader was also adopted.
Mass Spectrometry
QTRAP Liquid chromatography-mass spectrometry
Liquid chromatography-mass spectrometry was used to measure the C4-HSL concentration as a reference for QS system inducing.
Confocal
Nikon A1R HD25
Used for the visualization and analysis of more detailed dynamics of biofilm degradation
Inverted Fluorescence Microscope
Olympus IX73
Used for the observation of the dynamic culturing biofilm in our hardware.
Microfluidics System
CellASIC® ONIX B04A-03 Microfluidic Bacteria Plate
Extra system used in our study: Microfluidic system
At first, we considered two systems to measure the biofilm in the flowing condition: one was the original hardware、, and another was microfluidic system which had a high accuracy and was often used by biofilm scientists. We discussed the ideas with the Anti-Biofilm Community, Jingtao Liu’s lab and Guoqiang Chen’s lab during the integrated HP process, confirming and improving our hardware designs. Then we developed our microfluidic software and installed it into the microfluidic system. However, due to the limited time and human resources, we haven’t cultivated the biofilm of PAO1-GFP in the microfluidic system. It becomes a work which needs our future exploration.
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