Pierre Joseph

Micropipette aspiration (MPA) is one of the gold standards for quantifying biological samples' mechanical properties, which are crucial from the cell membrane scale to the multicellular tissue. However, relying on the manipulation of individual home-made glass pipettes, MPA suffers from low throughput and no automation. Here, we introduce the sliding insert micropipette aspiration method, which permits parallelization and automation, thanks to the insertion of tubular pipettes, obtained by photolithography, within microfluidic channels. We show its application both at the lipid bilayer level, by probing vesicles to measure membrane bending and stretching moduli, and at the tissue level by quantifying the viscoelasticity of 3D cell aggregates. This approach opens the way to high-throughput, quantitative mechanical testing of many types of biological samples, from vesicles and individual cells to cell aggregates and explants, under dynamic physico-chemical stimuli.

Micropipette aspiration (MPA) is one of the gold standards to quantify biological samples’ mechanical properties, which are crucial from the cell membrane scale to the multicellular tissue. However, relying on the manipulation of individual home-made glass pipettes, MPA suffers from low throughput and difficult automation. Here, we introduce the sliding insert micropipette aspiration (SIMPA) method, that permits parallelization and automation, thanks to the insertion of tubular pipettes, obtained by photolithography, within microfluidic channels. We show its application both at the lipid bilayer level, by probing vesicles to measure membrane bending and stretching moduli, and at the tissue level by quantifying the viscoelasticity of 3D cell aggregates. This approach opens the way to high-throughput, quantitative mechanical testing of many types of biological samples, from vesicles and individual cells to cell aggregates and explants, under dynamic physico-chemical stimuli.

Speeding up and enhancing the performances of nucleic acid biosensing technologies have remained drivers for innovation. Here, we optimize a fluorimetry-based technology for DNA detection based on the concentration of linear targets paired with probes. The concentration module consists of a microfluidic channel with the shape of a funnel in which we monitor a viscoelastic flow and a counter-electrophoretic force. We report that the technology performs better with a target longer than 100 nucleotides (nt) and a probe shorter than 30 nt. We also prove that the control of the funnel geometry in 2.5D using grayscale lithography enhances sensitivity by 100-fold in comparison to chips obtained by conventional photolithography. With these optimized settings, we demonstrate a limit of detection of 4 fM in 30 s and a detection range of more than five decades. This technology hence provides an excellent balance between sensitivity and time to result.

We present µLAS, a lab-on-chip system that concentrates, separates, and detects DNA fragments in a single module. µLAS speeds up DNA size analysis in minutes using femtomolar amounts of amplified DNA. Here we tested the relevance of µLAS for sizing expanded trinucleotide repeats, which cause over 20 different neurological and neuromuscular disorders. Because the length of trinucleotide repeats correlates with the severity of the diseases, it is crucial to be able to size repeat tract length accurately and efficiently. Expanded trinucleotide repeats are however genetically unstable and difficult to amplify. Thus, the amount of amplified material to work with is often limited, making its analysis labor-intensive. We report the detection of heterogeneous allele lengths in 8 samples from myotonic dystrophy type 1 and Huntington disease patients with up to 750 CAG/CTG repeats in five minutes or less. The high sensitivity of the method allowed us to minimize the number of amplification cycles and thus reduce amplification artefacts without compromising the detection of the expanded allele. These results suggest that µLAS can speed up routine molecular biology applications of repetitive sequences and may improve the molecular diagnostic of expanded repeat disorders.

Blockage of pores by particles is found in many processes, including filtration and oil extraction. We present filtration experiments through a linear array of ten channels with one dimension which is sub-micron, through which a dilute dispersion of Brownian polystyrene spheres flows under the action of a fixed pressure drop. The growth rate of a clog formed by particles at a pore entrance systematically increases with the number of already saturated (entirely clogged) pores, indicating that there is an interaction or “cross-talk” between the pores. This observation is interpreted based on a phenomenological model, stating that a diffusive redistribution of particles occurs along the membrane, from clogged to free pores. This one-dimensional model could be extended to two-dimensional membranes.