Protocol for measuring the responses of multiple budding yeast strains to extracellular change in parallel using a microfluidic device

Alcatras microfluidic devices allow long-term, single-cell measurements by immobilizing cells of budding yeast in hundreds of individual traps while removing daughters by the flow of a medium. Here, we present a protocol for simultaneously monitoring the growth and responses of up to five budding yeast strains using a multi-chamber version of the device. We describe steps for preparing logarithmic cultures and medium reservoirs, filling the device, and loading cells. We then detail procedures for securing tubing and starting the medium flow. For complete details on the use and execution of this protocol, please refer to Granados et al.¹.