The present invention relates to a particle arrangement transportation device and a particle arrangement transportation method for generating and transporting an arrangement of particles such as bacterial cells.
Techniques for quantitating bacterial cells have a wide range of applications not only in a fundamental research area of biotechnology but also in the fields of medicine, food, hygienic management, and the like (refer to NPL 1). For example, in medical practice, a physical condition of a patient is managed by quantitating bacterial cells contained in skin, mucous membrane, or urine of the patient. In the food sector, a bacterial count is monitored in order to obtain a control guideline of food fermentation. In this manner, a portable bacterial counter that supports on-site measurement is required in various fields.
Conventionally, as methods of quantitating bacterial cells, methods which utilize image recognition such as a colony method and fluorochrome staining and flow cytometry techniques which optically analyze individual bacterial cells have been proposed (refer to NPL 2).
However, the colony method and fluorochrome staining are methods that require cultivation of bacteria while flow cytometry techniques are optical methods and therefore require large-sized apparatuses. Therefore, the methods disclosed in NPL 2 all lack portability.
On the other hand, a recently proposed method analyzes a pattern of electrical pulses flowing through gold micropores embedded with peptides to quantitate bacterial cells that pass through the micropores (refer to NPL 3). According to the method disclosed in NPL 3, a portable bacterial counter capable of detecting individual bacterial cells can be realized.
However, since the method disclosed in NPL 3 requires generating an arrangement of bacterial cells and transporting the arrangement to the micropores in order to detect individual bacterial cells, there is a problem in that an apparatus capable of generating and transporting such an arrangement of bacterial cells is yet to be realized.
[NPL 1] O. Lazcka, et al., “Pathogen detection: A perspective of traditional methods and biosensors”, Biosensors and Bioelectronics, Vol. 22, pp. 1205-1217, 2007
[NPL 2] R. Hazan, et al., “A method for high throughput determination of viable bacteria cell counts in 96-well plates”, BMC Microbiology, Vol. 12, No. 259, 2012
[NPL 3] M. Tsutsui, et al., “Identification of Individual Bacterial Cells through the Intermolecular Interactions with Peptide-Functionalized Solid-State Pores”, Analytical Chemistry, Vol. 90, pp. 1511-1515, 2018.
Embodiments of the present invention have been made in order to solve the problem described above and an object thereof is to provide a particle arrangement transportation device and a particle arrangement transportation method capable of generating an arrangement of particles such as bacterial cells and transporting the arrangement to a specific location.
A particle arrangement transportation device according to embodiments of the present invention includes: a base material in which is formed a flow channel from an inlet port-side opening through which a solution containing particles to be an object of arrangement and transportation is introduced to an outlet port-side opening; a first electrode formed along the flow channel on a wall surface of the base material being exposed in the flow channel; second and third electrodes formed along the flow channel in the base material on both sides of the flow channel; and a power supply configured to apply an AC voltage between the first electrode and the second electrode and between the first electrode and the third electrode.
In addition, a particle arrangement transportation method according to embodiments of the present invention includes, with respect to a particle arrangement transportation device including a base material in which is formed a flow channel from an inlet port-side opening to an outlet port-side opening, a first electrode formed along the flow channel on a wall surface of the base material being exposed in the flow channel, and second and third electrodes formed along the flow channel in the base material on both sides of the flow channel: a first step of applying an AC voltage between the first electrode and the second electrode and between the first electrode and the third electrode; and a second step of introducing a solution containing particles to be an object of arrangement and transportation into the flow channel through the inlet port-side opening.
According to embodiments of the present invention, by providing a base material in which a flow channel is formed, a first electrode formed along the flow channel on a wall surface of the base material being exposed in the flow channel, second and third electrodes formed along the flow channel in the base material on both sides of the flow channel, and a power supply that applies an AC voltage between the electrodes, an arrangement of particles such as bacterial cells can be generated and transported to a specific location.
Hereinafter, an embodiment of the present invention will be described with reference to the drawings.
The base material 2 is constituted of a plate-like substrate 2a and a plate-like substrate 2b to be bonded to the substrate 2a. The electrodes 6 to 8 are formed on an upper surface of the substrate 2a. The groove-like flow channel 5 is formed on a lower surface of the substrate 2b at a position where the flow channel 5 covers the electrode 6 when the substrate 2a and the substrate 2b are bonded to each other by removing a lower surface side of the substrate 2b so as to retain an upper surface side thereof. A width W and a height H of the flow channel 5 must be set to sufficiently large values with respect to particles to be an object of arrangement and transportation. A length of the flow channel 5 can be arbitrarily set.
As a material of the substrate 2a and the substrate 2b, materials such as glass, silicon, and plastic can be used.
The substrate 2a and the substrate 2b are bonded so that the lower surface of the substrate 2b comes into contact with the upper surface of the substrate 2a, the flow channel 5 covers the electrode 6, and a lid of the flow channel 5 is closed.
The power supply 9 applies an AC voltage between the electrode 6 and the electrode 7 and between the electrode 6 and the electrode 8 (step S100 in
In addition, a pump (to be described later) feeds a solution 100 to the inlet port-side opening 3 of the particle arrangement transportation device 1 via a pipe connected to the inlet port-side opening 3 (step S101 in
In expression (1), εs denotes a complex dielectric constant of the solution 100, εp denotes a complex dielectric constant of the particles 101, r denotes a radius of the particles 101, E denotes electrical field intensity, and ∇ denotes a nabla operator. Re [] signifies a real part of a complex number described in []. The dielectrophoretic force F is described in reference literature "K. Mogi, et al., “Trapping and isolation of single prokaryotic cells in a micro-chamber array using dielectrophoresis”, RSC Advances, Vol. 6, pp.113000-113006, 2016”.
Since the complex dielectric constants εs and εp are dependent on a frequency of the AC voltage applied to the electrodes 6 to 8, a direction of the dielectrophoretic force F is also dependent on the frequency of the AC voltage. In the present embodiment, applying an AC voltage with a frequency satisfying εs < εp to the electrodes 6 to 8 enables the particles 101 to be fixed near the electrode 6 which is a high-electric field region in the flow channel 5.
When considering a case where the electrode 6 has a linear shape, since the lines of electric force L1 and L2 when the AC voltage is applied between the electrode 6 and the electrode 7 and between the electrode 6 and the electrode 8 concentrate at edges on both sides of the electrode 6 as shown in
In addition, by increasing the width W of the electrode 6 to twice the diameter of the particles 101 or more, an arrangement in which the particles 101 more or less line up in two rows along the edges on both sides of the electrode 6 can be generated as shown in
In this manner, by adjusting a disposition or a size of the electrode 6, an arbitrary arrangement of the particles 101 can be generated.
Generating a flow of the solution 100 enables the arranged particles 101 to be transported in a flow direction of the solution.
As a method of generating the flow of the solution 100, as shown in
However, the use of the pump 10 is not an essential constituent element in the present invention. The particle arrangement transportation device 1 may be installed so that the inlet port-side opening 3 is above and the outlet port-side opening 4 is below in order to dispose the flow channel 5 vertically downward or diagonally downward.
As described above, in the present embodiment, an arrangement of particles such as bacterial cells can be generated and transported to a specific location.
Combining the particle arrangement transportation device 1 according to the present embodiment with the sensor disclosed in NPL 3 and causing the particle arrangement transportation device 1 to transport bacterial cells to the micropores of the sensor enables the bacterial cells to be quantitated.
While the present embodiment has been described using bacterial cells as an example of particles, it is needless to say that the present invention can also be applied to particles other than bacterial cells.
Embodiments of the present invention can be applied to techniques for transporting particles.
1 Particle arrangement transportation device
2 Base material
5 Flow channel
6, 7, 8 Electrode
9 Power supply
10 Pump.
This application is a national phase entry of PCT Application No. PCT/JP2020/004855, filed on Feb. 7, 2020, which application is hereby incorporated herein by reference.
Filing Document | Filing Date | Country | Kind |
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PCT/JP2020/004855 | 2/7/2020 | WO |