High-throughput optical screening of cellular mechanotransduction
نویسندگان
چکیده
We introduce an optical platform for rapid, high-throughput screening of exogenous molecules that affect cellular mechanotransduction. Our method initiates mechanotransduction in adherent cells using single laser-microbeam generated micro-cavitation bubbles (μCBs) without requiring flow chambers or microfluidics. These μCBs expose adherent cells to a microTsunami, a transient microscale burst of hydrodynamic shear stress, which stimulates cells over areas approaching 1mm2. We demonstrate microTsunami-initiated mechanosignalling in primary human endothelial cells. This observed signalling is consistent with G-protein-coupled receptor stimulation resulting in Ca2+ release by the endoplasmic reticulum. Moreover, we demonstrate the dose-dependent modulation of microTsunami-induced Ca2+ signalling by introducing a known inhibitor to this pathway. The imaging of Ca2+ signalling, and its modulation by exogenous molecules, demonstrates the capacity to initiate and assess cellular mechanosignalling in real-time. We utilize this capability to screen the effects of a set of small molecules on cellular mechanotransduction in 96-well plates using standard imaging cytometry.
منابع مشابه
Nootropic Medicinal Plants; Evaluating Potent Formulation By Novelestic High throughput Pharmacological Screening (HTPS) Method
The principle of this method was to screen the pharmacological activity of six prepared polyphyto formulations by using high throughput screening method for their nootropic action. The study was performed in three stages using one, two and three animals, respectively in a group. Test formulations were given p.o daily at the dose of 50 and 100 mg/kg body weight. The test formulations were compar...
متن کاملHigh-throughput Cellular-resolution in Vivo Vertebrate Screening
Due to its small size and optical transparency, zebrafish larvae are excellent vertebrate models to study human diseases in vivo. We previously developed a high throughput screening platform capable of handling, cellular-resolution imaging and optically manipulating zebrafish larvae. Here, we present new novel technologies to significantly increase the throughput of our screening platform by mu...
متن کاملCell-Based Assays in High-Throughput Screening for Drug Discovery
Drug screening is a long and costly process confronted with low productivity and challenges in using animals, which limit the discovery of new drugs. To improve drug screening efficacy and minimize animal testing, recent efforts have been dedicated to developing cell-based high throughput screening (HTS) platforms that can provide more relevant in vivo biological information than biochemical as...
متن کاملTwo-photon photoactivated voltage imaging in tissue with an Archaerhodopsin-derived reporter
Robust voltage imaging in tissue remains a technical challenge. Existing combinations of genetically encoded voltage indicators (GEVIs) and microscopy techniques cannot simultaneously achieve sufficiently high voltage sensitivity, background rejection, and time resolution for highresolution mapping of sub-cellular voltage dynamics in intact brain tissue. We developed a pooled high-throughput sc...
متن کاملAdd-on for High Throughput Screening in Material Discovery for Organic Electronics: “Tagging” Molecules to Address the Device Considerations
This work reflects the worth of intelligent modeling in controlling the nanostructure morphology in manufacturing organic bulk heterojunction (BHJ) solar cells. It suggests the idea of screening the pool of material design possibilities inspired by machine learning. To fulfill this goal, a set of experimental data on a BHJ solar cell with a donor structure of diketopyrrolopyrrole (DDP) and ...
متن کامل