Development of micro‐inclined well array for trapping single cells

Tsukamoto, Takuya; Furuya, Naoki; Shimagami, Takuya; Sato, Rui; Shimokawa, Fusao; Akimitsu, Kazuya; Suzuki, Takao

doi:10.1002/ecj.12117

Cited by 3 publications

(4 citation statements)

References 16 publications

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“…29) After SU-8 cooling, the PDMS mold and the substrate are separated. After that, UV light is exposed with a normal exposure dose for an SU-8 submillimeter thickness from the back side of the Cr-patterned glass substrate 30) [Fig. 3(i)].…”

Section: Fabrication Processmentioning

confidence: 99%

Seamless fabrication technique for micro to millimeter structures by combining 3D printing and photolithography

Tamura

Suzuki

2019

Jpn. J. Appl. Phys.

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This paper proposes a seamless fabrication technique for micro-electro-mechanical systems (MEMS) devices having structures from micrometer to millimeter with a single material by combining 3D printing and photolithography. The proposed fabrication technique combines the fabrication characteristics of 3D printing and photolithography. We fabricated a mold of a microfluidic device having micro-and millimeter structures and evaluated three characteristics of the proposed technique. From the evaluations, the micro-to millimeter structures were seamlessly fabricated with an error within ±5%. The rough surface of the resist transferred from a 3D-printed master mold causes irregular reflection of UV light and reduction in patterning accuracy. The relation of object shape and fabrication of millimeter structures was evaluated. In addition, various advantages of the proposed fabrication technique were confirmed by demonstrating a fluid transportation test for the fabricated microfludic device.

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Section: Fabrication Processmentioning

confidence: 99%

Seamless fabrication technique for micro to millimeter structures by combining 3D printing and photolithography

Tamura

Suzuki

2019

Jpn. J. Appl. Phys.

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“…The two-dimensional geometry of a conventional well fabricated using photolithography is flexible in design [ 29 , 30 , 31 ]. In contrast, we propose an SCM with inverse-tapered three-dimensional (3D) wells (fabricated via 3D photolithography) to achieve single-cell immobilization and prevent cell dropout during the liquid exchange [ 32 ]. We carried out immobilization experiments with protoplasts of single plant cells with a mean diameter of 30 μm and found that the inverse-tapered wells contributed to a higher cell residual rate after one wash than the vertical wells.…”

Section: Introductionmentioning

confidence: 99%

Single-Cell Microarray Chip with Inverse-Tapered Wells to Maintain High Ratio of Cell Trapping

et al. 2023

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A single-cell microarray (SCM) influenced by gravitational force is expected to be one of the simple methods in various fields such as DNA analysis and antibody production. After trapping the cells in the SCM chip, it is necessary to remove the liquid from the SCM to wash away the un-trapped cells on the chip and treat the reagents for analysis. The flow generated during this liquid exchange causes the trapped cells to drop out of conventional vertical wells. In this study, we propose an inverse-tapered well to keep trapped cells from escaping from the SCM. The wells with tapered side walls have a reduced force of flow toward the opening, which prevents trapped cells from escaping. The proposed SCM chip was fabricated using 3D photolithography and polydimethylsiloxane molding techniques. In the trapping experiment using HeLa cells, the cell residual rate increased more than two-fold for the SCM chip with the inverse-tapered well with a taper angle of 30° compared to that for the conventional vertical SCM chip after multiple rounds of liquid exchanges. The proposed well structure increases the number of trapped cells and decreases the cell dropout rate to improve the efficiency of cellular analysis.

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“…3−8 Among these techniques, microfluidic arrays have been most commonly applied for their ability to simultaneously immobilize many single cells and enable in situ observation over time. 9 However, their complicated fabrication and the associated expense of photolithography and soft lithography have hindered the commercialization of these chips. 10,11 In addition, most of the trapped cells are always kept in round traps, which might influence their cellular functions.…”

Section: Introductionmentioning

confidence: 99%

“…In the analysis of single cells, to prevent interference from cell–cell interaction, trapping or immobilizing individual cells is a fundamental task. Numerous techniques have been developed to create microarrays that can trap single cells, based on the guidance of optical, magnetic, electrical, centrifugation, ultrasonic, pressure, and hydrodynamic forces. − Among these techniques, microfluidic arrays have been most commonly applied for their ability to simultaneously immobilize many single cells and enable in situ observation over time . However, their complicated fabrication and the associated expense of photolithography and soft lithography have hindered the commercialization of these chips. , In addition, most of the trapped cells are always kept in round traps, which might influence their cellular functions.…”

Section: Introductionmentioning

confidence: 99%

Single Polar Cell Trapping Based on the Breath Figure Method

Liu

Gao

et al. 2019

ACS Omega

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The ability to research individual cells is important for various biological studies. Currently reported biointerfaces for single-cell analysis can only trap individual cells in random morphologies. Cell polarity is a key factor in cellular functions, and the study of single-cell polarity can facilitate an understanding of cancer metastasis and stem-cell differentiation. For single polar cell trapping, anisotropic honeycomb-structured films were prepared. Elastic poly(1,2-butadiene) honeycomb films with ordered hexagonal pores were first prepared via the breath figure method. Subsequently, the films were subjected to mechanical stretching and fixed via photo-cross-linking under UV light irradiation. This stretched honeycomb structure was then transferred to a polystyrene surface. The resultant anisotropic porous films exhibited excellent capacity for single-cell trapping. Besides contributing to the physical spatial confinement of cells, the trapped single cells exhibited orientation in different polarities. The single polar cell array provided a novel platform for fundamental biological research.

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Development of micro‐inclined well array for trapping single cells

Cited by 3 publications

References 16 publications

Seamless fabrication technique for micro to millimeter structures by combining 3D printing and photolithography

Seamless fabrication technique for micro to millimeter structures by combining 3D printing and photolithography

Single-Cell Microarray Chip with Inverse-Tapered Wells to Maintain High Ratio of Cell Trapping

Single Polar Cell Trapping Based on the Breath Figure Method

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