Robotic Cell Printing for Constructing Living Yeast Cell Microarrays in Microfluidic Chips

Abstract

Living cell microarrays in microfluidic chips allow the non-invasive multiplexed molecular analysis of single cells. Here, we developed a simple and affordable perfusion microfluidic chip containing a living yeast cell array composed of a population of cell variants (green fluorescent protein (GFP)-tagged Saccharomyces cerevisiae clones). We combined mechanical patterning in 102 microwells and robotic piezoelectric cell dispensing in the microwells to construct the cell arrays. Robotic yeast cell dispensing of a yeast collection from a multiwell plate to the microfluidic chip microwells was optimized. The developed microfluidic chip and procedure were validated by observing the growth of GFP-tagged yeast clones that are linked to the cell cycle by time-lapse fluorescence microscopy over a few generations. The developed microfluidic technology has the potential to be easily upscaled to a high-density cell array allowing us to perform dynamic proteomics and localizomics experiments.

Development of bone cell microarrays in microfluidic chips for studying osteocyte-osteoblast communication under fluid flow mechanical loading

“…Moreover, the M2MD microdispenser is mounted with a conventional 10 µl plastic tip (epT.I.P.S. LoRetention from Eppendorf), which nozzle is significantly larger than the one of a piezoelectric glass tip for picolitre volume dispensing [39,62]. Although the printing resolution is reduced, larger nozzles are beneficial for decreasing the shear stress during the printing of sensitive cells [39,43].…”

“…LoRetention from Eppendorf), which nozzle is significantly larger than the one of a piezoelectric glass tip for picolitre volume dispensing [39,62]. Although the printing resolution is reduced, larger nozzles are beneficial for decreasing the shear stress during the printing of sensitive cells [39,43]. The use of plastic tips is also a more time-and cost-effective solution compared to the expensive custom-made glass tips, which can easily break and require extensive cleaning before reliable shooting.…”

“…However, robotic spotters allow for many other designs to be printed by simply adapting the x-y coordinates of each spot (e.g. words, flower, yin and yang, smileys, …) [39,62]. We took advantage of this flexibility to construct innovative arrays of osteoblast and osteocyte spots making contact with each other at precise interfaces (figure 3).…”

“…This was notably possible due to recent developments in microfluidic technology, giving rise to miniaturised and integrated devices suitable to perform intercellular communication experiments on-chips [31][32][33][34][35][36]. Contrarily to multiwell plate experiments, microfluidic chips enable to dynamically control the cell microenvironment by perfusing precise volumes of cell medium supplemented with nutrients, drugs, metabolites, etc [37][38][39][40]. The perfusion rate through microfluidic channels can be easily adapted, which is useful to perform e.g.…”

Yeast Biotechnology 3.0

“…Living cell microarrays in microfluidic chips allow the non-invasive multiplexed molecular analysis of single cells. Yvanoff et al [1] developed a simple and affordable perfusion microfluidic chip containing a living yeast cell array composed of a population of green fluorescent protein (GFP)-tagged Saccharomyces cerevisiae clones. Mechanical patterning in microwells and robotic piezoelectric cell dispensing in the microwells were combined to construct the cell arrays.…”