A microfluidic device for studying multiple distinct strains

J Vis Exp. 2012 Nov 9:(69):4257. doi: 10.3791/4257.

Abstract

The study of cell responses to environmental changes poses many experimental challenges: cells need to be imaged under changing conditions, often in a comparative manner. Multiwell plates are routinely used to compare many different strains or cell lines, but allow limited control over the environment dynamics. Microfluidic devices, on the other hand, allow exquisite dynamic control over the surrounding conditions, but it is challenging to image and distinguish more than a few strains in them. Here we describe a method to easily and rapidly manufacture a microfluidic device capable of applying dynamically changing conditions to multiple distinct yeast strains in one channel. The device is designed and manufactured by simple means without the need for soft lithography. It is composed of a Y-shaped flow channel attached to a second layer harboring microwells. The strains are placed in separate microwells, and imaged under the exact same dynamic conditions. We demonstrate the use of the device for measuring protein localization responses to pulses of nutrient changes in different yeast strains.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Concanavalin A / chemistry
  • Dimethylpolysiloxanes / chemistry
  • Glass / chemistry
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods*
  • Nylons / chemistry
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae / classification*
  • Saccharomyces cerevisiae / cytology

Substances

  • Dimethylpolysiloxanes
  • Nylons
  • poly(dimethylsiloxane)-polyamide copolymer
  • Concanavalin A