A method for screening microorganisms using supramolecular fluorescence microfluidic technology
A microfluidic technology, a technology of supramolecular complexes, is applied in the field of high-throughput screening of microorganisms realized by supramolecular fluorescence microfluidics, which can solve the problems of inability to detect metabolites, complex labeling process, and limited application scope, and achieve The effect of high detection sensitivity, low detection cost and simple experimental operation
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Embodiment 1
[0025] Analysis of tryptophan acid concentration within microfluidic droplets using supramolecular fluorescence.
[0026] Different concentrations of tryptophan were added to the supramolecular fluorescent supramolecular solution of CB[8] and MDPP with a molar ratio of 1:1, and the fluorescence intensity of the supramolecular solution was detected by a microplate reader. attached by Figure 5 It can be seen that when tryptophan is added to the supramolecular fluorescence system, the fluorescence of the system gradually decreases with the addition of tryptophan, which is because tryptophan enters the remaining cavity of CB8, and the aromatic group of tryptophan and MDPP π-π stacking is generated, resulting in fluorescence quenching.
[0027] Three groups of solutions were prepared: water phase (1): CB[8] (5μM) and MDPP (5μM); water phase (2): CB[8] (5μM), MDPP (5μM) and tryptophan (5μM) ; Aqueous phase (3): CB[8] (5 μM), MDPP (5 μM) and tryptophan (10 μM). The flow rate of t...
Embodiment 2
[0029] Supramolecular fluorescence analysis of tryptophan concentrations metabolized by Escherichia coli within microfluidic droplets.
[0030] The E. coli suspension was diluted to 106 / mL with liquid medium. The basic components of the medium are: trace elements 0.1%, K 2 HPO 4 2.5%, KH 2 PO 4 0.9%, (NH 4 ) 2 SO 4 0.45%, glucose 0.3%, MgSO 0.01% pH7.2. , select the cell wrapping chip for cell droplet wrapping, set the flow rate of the oil phase to 3 μL / min, and the flow rate of the E. coli cell diluent to 1 μL / min to generate droplets into a 1 mL syringe with a diameter of 30 μm. According to the Poisson distribution , the proportion of droplets without cells is 0.6065, the proportion of droplets embedded with single cells is 0.3033, and the proportion of droplets embedded with two or more cells is 0.0902. The syringe was placed in a 37°C incubator for 24 hours, and the cucurbituril / fluorescent dye supramolecular complex was injected into the droplets by microfluidic picolit...
Embodiment 3
[0032] Screening of Escherichia coli with different tryptophan metabolism ability using supramolecular fluorescence microfluidic technology.
[0033]Place the droplet detection chip on the stage, and adjust the position of the stage so that the droplet detection chip can be clearly imaged on the high-speed camera. Set the droplet flow rate to 0.1 μL / min and the oil phase flow rate to 2 μL / min, inject the droplets into the droplet detection chip, turn on the laser, set the laser wavelength to 442nm, and the detection wavelength to 500-520nm, and adjust the droplet detection chip Position the laser at the channel detection point of the drop detection chip. Turn on the photomultiplier tube, set the trigger value of the signal analysis and processing software, and the gain value of the photomultiplier tube. The waveform graph displays the trigger threshold value and the droplet fluorescence intensity value. When the detection value of the detected target droplet exceeds the predet...
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