Category: Automation and High-Throughput Technologies
The advent of induced pluripotent stem cells combined with advances in organ-on-a-chip technologies has led to the development of microfluidic neuronal and glial models. The rapid advances in the field of 3D cell culture and Organ on a Chip models for drug screening and toxicity studies are creating a demand for electrophysiological readout technology development.
In conventional models, electrophysiological characterization is commonly perform using patch clamping methods. While morphological characterization using immunohistochemistry is easily transferred into a microfluidic setting, electrophysiological characterization remains underdeveloped. Calcium imaging has been used to assess electrophysiology without the need for direct contact with electrodes, but the methods do not yet match the high throughput potential offered by microfluidic systems. To this end, we developed a solution to evaluate the electrophysiological activity of neuronal models in a commercially available Organ-on-a-Chip platform, the OrganoPlate®. The OrganoPlate is a microfluidic platform enabling the culture of 96 individual three-dimensional organ models in a format compatible with standard screening platforms.
Our method enables high throughput compatible analysis of iPSCs derived neuron and glia in the Mimetas OrganoPlate. Using a Molecular Devices ImageXpress Micro XLS-C high a content microscope recording at 2 Hz, we are able to extract and quantify the Electrophysiological activity of neuronal models in the OrganoPlate. This is a data conscious departure from high frequency based methods employed in recent literature, where gigabytes large streams are acquired for a single chip. Our method is capable of automatic extraction and analysis of each individual neuron in the neuronal network. The ImageJ based software solution is capable of measuring and quantifying individual neuronal spike trains, providing statistical information of the entire network within two minutes per chip. The low framerate of acquisition combined with fluorescent imaging permits the use of calcium imaging using widely available microscopy equipment and computing resources. Overall, the combination a high throughput microfluidic 3D cell culture platform, high content imaging and automated image analysis provides a valuable tool for assessing the effects of drug candidates on neuronal firing.
Arnaud Nicolas– Junior Scientist, Leiden University / Mimetas, Leiden, Zuid-Holland, Netherlands
Leiden University / Mimetas
Leiden, Zuid-Holland, Netherlands
With a Master in micro and nano electronics and an engineering degree in biomedical sciences, Arnaud has set himself to pursue a PhD in the innovative field of biomedical sciences and Organ-on-a-Chip technologies. During three years as a graduate student from the Leiden University, Arnaud has been working with Mimetas, a science based startup, to develop innovative assays and applications compatible with a wide array of laboratory platforms, including microscopy, confocal microscopy, TEER measurements and Calcium imaging. The common denominator of the projects he engages in? A sincere collaboration with programmers, engineers and biologists to bring forward tomorrow's biomedical advances.