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

Charlotte Yvanoff,Ronnie G Willaert,Stefania Torino

doi:10.3390/fermentation6010026

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.

Highlights

Cell assays have been miniaturized by growing cells in multiwell plates with increasing well number from 96 to 384 and 1536 wells and decreasing well volume from 280 μL down to 3 μL
We developed a perfusion microfluidic chip containing living yeast cell arrays that allows long term cultivation of the yeast cells and high-resolution time-lapse fluorescence microscopy
The creation of the cell array was based on mechanical patterning in SU-8 microwells and piezoelectric filling of the microwells

Summary

Introduction

Cell assays have been miniaturized by growing cells in multiwell plates with increasing well number from 96 to 384 and 1536 wells and decreasing well volume from 280 μL down to 3 μL. These experiments are typically integrated in a robotic analysis platform. Cell microarrays provide an attractive solution, as they could increase the throughput significantly [1,2,3]. Living cell microarrays have been combined with microfluidic bioreactors, which provide multiple advantages for multiplex dynamic analyses and high-throughput screening [4,6]. An interesting application where living yeast cell arrays are used is “dynamic proteomics”

Methods

Results

Discussion

Conclusion