Misawa N et al. (2023), Xenopus laevis Oocyte Array Fluidic Device Inte...

XB-ART-59574

Sensors (Basel) 2023 Feb 21;235:. doi: 10.3390/s23052370.

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Xenopus laevis Oocyte Array Fluidic Device Integrated with Microelectrodes for A Compact Two-Electrode Voltage Clamping System.

Misawa N , Tomida M , Murakami Y , Mitsuno H , Kanzaki R .

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We report on a compact two-electrode voltage clamping system composed of microfabricated electrodes and a fluidic device for Xenopus laevis oocytes. The device was fabricated by assembling Si-based electrode chips and acrylic frames to form fluidic channels. After the installation of Xenopus oocytes into the fluidic channels, the device can be separated in order to measure changes in oocyte plasma membrane potential in each channel using an external amplifier. Using fluid simulations and experiments, we investigated the success rates of Xenopus oocyte arrays and electrode insertion with respect to the flow rate. We successfully located each oocyte in the array and detected oocyte responses to chemical stimuli using our device.

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Species referenced: Xenopus laevis

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References [+] :

Arakawa, A highly sensitive and temporal visualization system for gaseous ethanol with chemiluminescence enhancer. 2012, Pubmed

	Figure 1. (a) Schematic images of the fluidic device with electrodes for an array of four oocytes. For surface insulating of the electrode chip, the chip was entirely covered with parylene-C, except for the contact pads and electrode tips (yellow areas). (b) Section images correspond to the colored dashed lines of a-a’, b-b’, c-c’, d-d’, and e-e’ in (a).
	Figure 2. (a) Image of the device. (b) A single device unit. (c) Micrograph image (top view) of the electrodes. (d) Scanning electron microscopic image of an electrode tip.
	Figure 3. Stream renderings for fluid simulations at an entry flow rate of (a) 1 mL min−1, (b) 5 mL min−1, (c) 10 mL min−1 before oocyte trapping, and (d) 10 mL min−1 after oocyte trapping. Partial top and side views are shown in point of sights “A” and “B”, indicated by arrows in the upper left panels (overhead views).
	Figure 4. Superimposed images of oocyte motion; (a) an oocyte bypassed at a flow rate of 3 mL min−1, and (b) four different oocytes trapped at a flow rate of 20 mL min−1. The position of each oocyte over time is indicated by an arrow. (c) The success rate of oocyte trapping with respect to flow rate. Each data point is more than eight experiments.
	Figure 5. Photos of an oocyte that is (a) not inserted (failure), (b) inserted (success), and (c) penetrated by electrodes or passed through the trap (failure). Flow direction was left to right in (a), (b), and (c). (d) Correlation between the flow rate and electrode invasion level. Each data point represents 12 observations per flow rate.
	Figure 6. The current trace of an oocyte expressing BmOR3. At the time points indicated by arrows, each operation (addition or removal of 20 µL solution) was carried out sequentially. (a) Barth’s solution without bombykal was added. (b) The solution was removed from the fluidic device. (c) 100 µM bombykal solution was added. (d) The solution was removed again.