Microfluidic Devices for Imaging and Biochemistry Analysis of Microbes Under Mechanical Pressure
- 1Laboratory for Analysis and Architecture of Systems, CNRS, Universite de Toulouse, Toulouse, France
Abstract
Growth-induced pressure arises when proliferating cell populations are confined within rigid microenvironments and is increasingly recognized as a determinant of microbial physiology in soils, biofilms, and host tissues. We present two complementary microfluidic devices that confine microbes within polydimethylsiloxane (PDMS) chambers and apply defined, optically read-out growth-induced pressure of up to 1.5 MPa. The first, the self-closing (SC) chip, confines cells in multiple small chambers accommodating hundreds of cells; it provides excellent nutrient supply and rapid medium exchange, supports single-cell imaging, and allows multiplexing of cellular or chemical conditions. The second, the pressure-recovery (PR) chip, confines cells in a single 5 cm-long chamber accommodating hundreds of thousands of cells; a scalpel-cut step recovers live cells from the channel within minutes for bulk biochemical assays. The two devices share a single two-layer soft-lithography fabrication process and a common brightfield wall-displacement pressure readout. Together, they enable single-cell imaging and bulk biochemical analysis under matched, defined pressure conditions. We illustrate the protocol with two representative validations: rapid {beta}-estradiol-induced transcription in the SC chip, in which nascent transcription foci appear within 5 min independently of the applied pressure, and the PR chip, which produces uniform growth-induced pressure along the entire 5 cm channel and recovers between 0.1x10^6 and 1x10^6 cells per device in a pressure-tunable manner.
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BibTeX
@misc{kim2026microfluidic,
title = {{Microfluidic Devices for Imaging and Biochemistry Analysis of Microbes Under Mechanical Pressure}},
author = {H. Kim and N. Nguyen and B. Alric and L. Albert and M. Delarue},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.06.749717},
url = {https://www.biorxiv.org/content/10.64898/2026.09.06.749717v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Microfluidic Devices for Imaging and Biochemistry Analysis of Microbes Under Mechanical Pressure
AU - H. Kim
AU - N. Nguyen
AU - B. Alric
AU - L. Albert
AU - M. Delarue
PY - 2026
DA - 2026/09/06
PB - bioRxiv
DO - 10.64898/2026.09.06.749717
UR - https://www.biorxiv.org/content/10.64898/2026.09.06.749717v1
AB - Growth-induced pressure arises when proliferating cell populations are confined within rigid microenvironments and is increasingly recognized as a determinant of microbial physiology in soils, biofilms, and host tissues. We present two complementary microfluidic devices that confine microbes within polydimethylsiloxane (PDMS) chambers and apply defined, optically read-out growth-induced pressure of up to 1.5 MPa. The first, the self-closing (SC) chip, confines cells in multiple small chambers accommodating hundreds of cells; it provides excellent nutrient supply and rapid medium exchange, supports single-cell imaging, and allows multiplexing of cellular or chemical conditions. The second, the pressure-recovery (PR) chip, confines cells in a single 5 cm-long chamber accommodating hundreds of thousands of cells; a scalpel-cut step recovers live cells from the channel within minutes for bulk biochemical assays. The two devices share a single two-layer soft-lithography fabrication process and a common brightfield wall-displacement pressure readout. Together, they enable single-cell imaging and bulk biochemical analysis under matched, defined pressure conditions. We illustrate the protocol with two representative validations: rapid {beta}-estradiol-induced transcription in the SC chip, in which nascent transcription foci appear within 5 min independently of the applied pressure, and the PR chip, which produces uniform growth-induced pressure along the entire 5 cm channel and recovers between 0.1x10^6 and 1x10^6 cells per device in a pressure-tunable manner.
ER -