Studying Glycolytic Oscillations in Individual Yeast Cells by Combining Fluorescence Microscopy with Microfluidics and Optical Tweezers

Anna Karin Gustavsson*, Amin A. Banaeiyan, David D. van Niekerk, Jacky L. Snoep, Caroline B. Adiels, Mattias Goksör

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

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.

Original languageEnglish
Article numbere70
Pages (from-to)1-26
Number of pages26
JournalCurrent Protocols in Cell Biology
Volume82
Issue number1
Early online date17 Oct 2018
DOIs
Publication statusPublished - Mar 2019

Keywords

  • fluorescence microscopy
  • glycolytic oscillations
  • microfluidics
  • optical tweezers
  • single-cell analysis

Fingerprint Dive into the research topics of 'Studying Glycolytic Oscillations in Individual Yeast Cells by Combining Fluorescence Microscopy with Microfluidics and Optical Tweezers'. Together they form a unique fingerprint.

Cite this