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

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.

Scaling of stochastic growth and division dynamics: A comparative study of individual rod-shaped cells in the Mother Machine and SChemostat platforms

“…Despite its numerous advantages, the Mother Machine has design constraints whose effects on qualitative and quantitative cell physiology merit closer examination. The loading procedure involves centrifugation of bacteria to force them into the dead-end channels, which may introduce stresses ( Tanaka et al. , 2023 ).…”

Scaling of stochastic growth and division dynamics: A comparative study of individual rod-shaped cells in the Mother Machine and SChemostat platforms

“…Despite its numerous advantages, the Mother Machine has design constraints whose effects on qualitative and quantitative cell physiology merit closer examination. The loading procedure involves centrifugation of bacteria to force them into the dead-end channels, which may introduce stresses (37). Most studies have used a dedicated fluorescent reporter to identify and segment cells (26), but overexpression of a fluorescent marker can introduce a growth burden (38) and prolonged exposure to requisite bright illumination can lead to phototoxicity (39).…”