Lab-on-chip microscope platform for electro-manipulation of a dense microtubules network

Daniel Havelka,Djamel Eddine Chafai,Michal Teplan,Ilia Zhernov,Zdeněk Lánský,Michal Cifra

doi:10.1038/s41598-022-06255-y

Abstract

Pulsed electric field (PEF) technology is promising for the manipulation of biomolecular components and has potential applications in biomedicine and bionanotechnology. Microtubules, nanoscopic tubular structures self-assembled from protein tubulin, serve as important components in basic cellular processes as well as in engineered biomolecular nanosystems. Recent studies in cell-based models have demonstrated that PEF affects the cytoskeleton, including microtubules. However, the direct effects of PEF on microtubules are not clear. In this work, we developed a lab-on-a-chip platform integrated with a total internal reflection fluorescence microscope system to elucidate the PEF effects on a microtubules network mimicking the cell-like density of microtubules. The designed platform enables the delivery of short (microsecond-scale), high-field-strength (le 25 kV/cm) electric pulses far from the electrode/electrolyte interface. We showed that microsecond PEF is capable of overcoming the non-covalent microtubule bonding force to the substrate and translocating the microtubules. This microsecond PEF effect combined with macromolecular crowding led to aggregation of microtubules. Our results expand the toolbox of bioelectronics technologies and electromagnetic tools for the manipulation of biomolecular nanoscopic systems and contribute to the understanding of microsecond PEF effects on a microtubule cytoskeleton.

Highlights

Pulsed electric field (PEF) technology is promising for the manipulation of biomolecular components and has potential applications in biomedicine and bionanotechnology
To fill in these technology and knowledge gaps, we present here the design, fabrication, and verification of a new chip assembly, embedded to a total internal reflection fluorescence (TIRF) microscope, that enables the delivery of μs-PEF
Chip microscopy platform that can be reproducibly fixed to a Nikon Eclipse Ti TIRF microscope stage and enables easy manipulation of the chip and delivery of μs-PEF to out-of-the-cell MT networks

Summary

Introduction

Pulsed electric field (PEF) technology is promising for the manipulation of biomolecular components and has potential applications in biomedicine and bionanotechnology. Several PEF chip-microscope integration technologies have been developed in recent years[25–27], none of them demonstrate the capability of μs-PEF delivery to out-of-the-cells MT networks in a microfluidic environment To fill in these technology and knowledge gaps, we present here the design, fabrication, and verification of a new chip assembly, embedded to a total internal reflection fluorescence (TIRF) microscope, that enables the delivery of μs-PEF. Using this new experimental platform, we demonstrate in vitro that μs-PEF can influence multivalent interactions between MTs and the substrate

Methods

Results

Conclusion