Schematic illustration (a), layout (b), and fabrication process (c) of the device for investigating the growth propagation of yeast in linear arrays of micro-chambers. Cells are first loaded into the up-most chambers and then grow sequentially along the linear arrays (blue). Nutrient is provided by diffusion from a multi-U shaped channel (red). The first level of the mold for soft lithography is defined by photolithography, lift-off, and reactive ion etching techniques on the surface of a silicon wafer. The second level is patterned by photolithography with a SU8 resist. After molding by soft lithography, the PDMS layer is irreversibly bonded to a cover glass.
Yeast growth in a liner array of 50 × 50 μm2 square chambers with 6 μm wide and 80 μm long connection channels. (a) DIC image of the last three occupied chambers. (b) Corresponding fluorescence image of MYO1-GFP protein. (c) Corresponding fluorescence image of MCM-mCherry protein.
Variation of the cell number as a function of culture time in one of the micro-chambers. Cells are sub-sequentially loaded into the chamber from a 6 μm wide and 40 μm long connection channel. The solid line is a fitting of the experimental data with a homogenous loading of 28 cells during 7 h. The dashed line is obtained with only one cell input.
Variation of the cell number as a function of culture time in a chamber with 12 μm wide and 20 μm long inter-connection channels. The solid line is a fitting with a homogeneous loading of 8 cells and two additional loading of 28 cells at 13 min and 30 cells at 75 min. The dashed line is obtained with a homogeneous cell loading after an initial loading of 28 cells at 13 min.
Variation of the number of occupied chambers as a function of culture time for linear arrays of the same chamber size but different geometries of connection channels.
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