Open AccessStudying Glycolytic Oscillations in Individual Yeast Cells by Combining Fluorescence Microscopy with Microfluidics and Optical Tweezers
Author(s) -
Gustavsson AnnaKarin,
Banaeiyan Amin A.,
Niekerk David D.,
Snoep Jacky L.,
Adiels Caroline B.,
Goksör Mattias
Publication year - 2019
Publication title -
current protocols in cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.149
H-Index - 38
eISSN - 1934-2616
pISSN - 1934-2500
DOI - 10.1002/cpcb.70
Subject(s) - optical tweezers , microfluidics , fluorescence microscope , microscopy , yeast , nanotechnology , fluorescence , biophysics , biology , chemistry , optics , biological system , materials science , physics , biochemistry
In this unit, we provide a clear exposition of the methodology employed to study dynamic responses in individual cells, using microfluidics for controlling and adjusting the cell environment, optical tweezers for precise cell positioning, and fluorescence microscopy for detecting intracellular responses. This unit focuses on the induction and study of glycolytic oscillations in single yeast cells, but the methodology can easily be adjusted to examine other biological questions and cell types. We present a step‐by‐step guide for fabrication of the microfluidic device, for alignment of the optical tweezers, for cell preparation, and for time‐lapse imaging of glycolytic oscillations in single cells, including a discussion of common pitfalls. A user who follows the protocols should be able to detect clear metabolite time traces over the course of up to an hour that are indicative of dynamics on the second scale in individual cells during fast and reversible environmental adjustments. © 2018 by John Wiley & Sons, Inc.