The transport, distribution, and mixing of microfluidics
often
require additional instruments, such as pumps and valves, which are
not feasible when operated in point-of-care (POC) settings. Here,
we present a simple microfluidic pathogen detection system known as
Rotation-Chip that transfers the reagents between wells by manually
rotating two concentric layers without using external instruments.
The Rotation-Chip is fabricated by a simple computer numerical control
(CNC) machining process and is capable of carrying out 60 multiplexed
reactions with a simple 30 or 60° rotation. Leveraging superhydrophobic
coating, a high fluid transport efficiency of 92.78% is achieved without
observable leaking. Integrated with an intracellular fluorescence
assay, an on-chip detection limit of 1.8 × 106 CFU/mL
is achieved for ampicillin-resistant Escherichia coli
(E. coli), which is similar to our off-chip results.
We also develop a computer vision method to automatically distinguish
positive and negative samples on the chip, showing 100% accuracy.
Our Rotation-Chip is simple, low-cost, high-throughput, and can display
test results with a single chip image, making it ideal for various
multiplexing POC applications in resource-limited settings.
The transport, distribution, and mixing of microfluidics often require additional instruments, such as pumps and valves, which are not feasible when operated in point-of-care (POC) settings. Here, we present a simple microfluidic pathogen detection system known as Rotation-Chip that transfers the reagents between wells by manually rotating two concentric layers without using external instruments. The Rotation-Chip is fabricated by a simple computer numerical control (CNC) machining process and is capable of carrying out 60 multiplexed reactions with a simple 30-degree or 60-degree rotation. Leveraging superhydrophobic coating, a high fluid transport efficiency of 92.78% is achieved without observable leaking. Integrated with an intracellular fluorescent assay, an on-chip detection limit of 1.8×106CFU/mL is achieved for ampicillin-resistantEscherichia coli (E. coli), which is similar to our off-chip results. We also develop a computer vision method to automatically distinguish positive and negative samples on the chip, showing 100% accuracy. Our Rotation-Chip is simple, low-cost, high-throughput, and can display test results with a single chip image, ideal for various multiplexing POC applications in resource-limited settings.
Previous studies identified that strigolactones (SLs) and gibberellins (GAs) interacted when controlling branching in plant shoots, but the underlying mechanism remains unknown. qRT-PCR analysis suggested that the SL receptor gene CsDAD2 was significantly upregulated in the leaves, stems, and nodes of cucumber after treatment with 50 mg/L of GA3. Furthermore, the CsDAD2 gene was cloned and introduced into wild-type Arabidopsis plants via Agrobacterium-mediated transformation. For the CsDAD2-OE lines, the endogenous content of GA3 was subsequently higher at the seedling stage, with the number of primary cauline branches also significantly increased at the maturity stage compared with WT. Additionally, GA-related genes were up-regulated in the first inter-nodes and the third nodes of the CsDAD2-OE lines, thus indicating that GA was metabolically active in these tissues. The expression of the branch inhibitor gene AtBRC1 decreased at the seedling stage as well as at the maturity stage of the CsDAD2-OE lines. These findings suggest that CsDAD2 might have important functions in the interactions between GAs and SLs as it can promote the accumulation of GAs in plant nodes and suppress the expression of BRC1, hence increasing primary cauline branching.
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