Abstract: Multiplexable microfluidic culture chamber for imaging monolayer growth of single cells The present invention relates generally to a microfluidic device (1a, 1b), particularly for use in single cell analysis. More specifically, the present invention relates to a microfluidic device (1a, 1b) comprising at least one chamber (10), in particular at least two chambers (10) comprising a deformable membrane (16) and having a structure and geometry configured to enable formation of two-dimensional cell culture, in particular two-dimensional cell growth area (29), and imaging thereof over a growth period or a time period sufficient to analyze cells, in particular to monitor cell growth. The microfluidic device (1a, 1b) allows for multi-condition operation of single-cell screening at high spatiotemporal resolution. The present invention also relates to methods for fabrication and use of such devices.