Fig. 4: MSBC platform built multispecies consortia with flexibility and precision.

a Faster-growing E. coli overtook the S. cerevisiae in a well-mixed consortium. Cell mixture (S. cerevisiae (BY4742 expressing GFP) and E. coli (MG1655 expressing mCherry) at different seeding ratios) were inoculated into the medium, cultured at 30 °C for 24 h and quantified by flow cytometry. The dotted and dashed lines showed the actual and ideal composition, respectively. The x axis indicated the seeding density of S. cerevisiae (\(\frac{{N}_{S.{cerevisiae}}}{{N}_{S.{cerevisiae}}+{N}_{E.{coli}}}\)). In all conditions, the faster-growing E. coli became dominant. b The growth of E. coli and S. cerevisiae in MSBC was constrained inside their own niche with a defined boundary. We mixed the MSB(S. cerevisiae) and MSB(E. coli) at different seeding ratios (~2000 MSBs in total), and cultured the MSBC at 30 °C for 24 h. In all conditions, two species grew inside their own MSBs without interference and were able to maintain a desired population ratio consequently. Numbers indicated the seeding density of MSB(E. coli) (\(\,\frac{{{MSB}}_{E.{coli}}}{{M{SB}}_{S.{cerevisiae}}+{{MSB}}_{E.{coli}}}\)). Scale bar = 400 μm. c MSBC enabled precise control in the composition. The dotted and dashed lines showed the actual and ideal composition, respectively (see “Methods”). Quantification results reflected that MSBC strategy could precisely modulate the composition of the consortia. The x axis indicated the seeding density of MSB(S. cerevisiae). Error bars = standard deviation (n = 3 biologically independent samples) in a, c. d The growth of various species were supported in MSB. We generated MSB (E. coli) (expressing BFP), MSB(S. cerevisiae)(expressing GFP) and MSB(C. glutamicum) (expressing RFP), respectively. All three species grew inside MSBs showing the desired fluorescence. e MSBC platform assembled multispecies consortia. We randomly assembled three two-species MSBC (chamber 1–3, seeding ratio = 1:1), and a three-species MSBC (chamber 4, seeding ratio = 1:1:1). In all chambers, multiple species co-existed and the consortia composition was precisely modulated by the seeding ratio. Scale bar = 1 mm. f Assembly of consortia can be customized by MSBC strategy. According to the designs (schematic on top), we were able to construct various MSBC by simply replacing the MSB of different species. Source data are provided as a Source Data file.