High-Efficiency Single-Cell Containment Microdevices Based on Fluid Control

Daiki Tanaka; Junichi Ishihara; Hiroki Takahashi; Masashi Kobayashi; Aya Miyazaki; Satsuki Kajiya; Risa Fujita; Naoki Maekawa; Yuriko Yamazaki; Akiko Takaya; Yuumi Nakamura; Masahiro Furuya; Tetsushi Sekiguchi; Shuichi Shoji

doi:10.3390/mi14051027

High-Efficiency Single-Cell Containment Microdevices Based on Fluid Control

Micromachines (Basel). 2023 May 11;14(5):1027. doi: 10.3390/mi14051027.

Authors

Daiki Tanaka¹, Junichi Ishihara², Hiroki Takahashi^{2

3

4}, Masashi Kobayashi⁵, Aya Miyazaki⁵, Satsuki Kajiya⁵, Risa Fujita¹, Naoki Maekawa⁶, Yuriko Yamazaki^{7

8

9}, Akiko Takaya^{2

4

6}, Yuumi Nakamura^{7

9}, Masahiro Furuya¹⁰, Tetsushi Sekiguchi¹, Shuichi Shoji⁵

Affiliations

¹ Research Organization for Nano & Life Innovation, Waseda University, 513 Waseda Tsurumakicho, Shinjuku-ku, Tokyo 162-0041, Japan.
² Medical Mycology Research Center, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-0856, Japan.
³ Molecular Chirality Research Center, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba 263-8522, Japan.
⁴ Plant Molecular Science Center, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8675, Japan.
⁵ School of Fundamental Science and Engineering, Waseda University, 3-4-1 Okubo, Shin-juku-ku, Tokyo 169-8555, Japan.
⁶ Department of Natural Products Chemistry, Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan.
⁷ Department of Dermatology, Osaka University Graduate School of Medicine, Osaka 565-0871, Japan.
⁸ Department of Dermatology, Chiba University Graduate School of Medicine, Chiba 260-8670, Japan.
⁹ Cutaneous Allergy and Host Defense, Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
¹⁰ Graduate School of Advanced Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, Japan.

Abstract

In this study, we developed a comb-shaped microfluidic device that can efficiently trap and culture a single cell (bacterium). Conventional culture devices have difficulty in trapping a single bacterium and often use a centrifuge to push the bacterium into the channel. The device developed in this study can store bacteria in almost all growth channels using the flowing fluid. In addition, chemical replacement can be performed in a few seconds, making this device suitable for culture experiments with resistant bacteria. The storage efficiency of microbeads that mimic bacteria was significantly improved from 0.2% to 84%. We used simulations to investigate the pressure loss in the growth channel. The pressure in the growth channel of the conventional device was more than 1400 PaG, whereas that of the new device was less than 400 PaG. Our microfluidic device was easily fabricated by a soft microelectromechanical systems method. The device was highly versatile and can be applied to various bacteria, such as Salmonella enterica serovar Typhimurium and Staphylococcus aureus.

Keywords: cell culture; fluid control; microfluidic devices; single cell.

Grants and funding

This work was supported by JST, ACT-X Grant Number JPMJAX21KA, Japan, and Japan Ministry of Education, Culture, Sports Science & Technology (MEXT) Grant-in-Aid for Scientific Basic Research (A) No. 20H00336, Grant-in-Aid for Young Scientists 17K15151 and 22K15152.