A Method for Imaging Oxygen Distribution and Respiration at a Microscopic Level of Resolution

Methods Mol Biol. 2017:1670:31-38. doi: 10.1007/978-1-4939-7292-0_3.

Abstract

Conventional oxygen (micro-) sensors assess oxygen concentration within a particular region or across a transect of tissue, but provide no information regarding its bidimensional distribution. Here, a novel imaging technology is presented, in which an optical sensor foil (i.e., the planar optode) is attached to the surface of the sample. The sensor converts a fluorescent signal into an oxygen value. Since each single image captures an entire area of the sample surface, the system is able to deduce the distribution of oxygen at a resolution level of few micrometers. It can be deployed to dynamically monitor oxygen consumption, thereby providing a detailed respiration map at close to cellular resolution. Here, we demonstrate the application of the imaging tool to developing plant seeds; the protocol is explained step by step and some potential pitfalls are discussed.

Keywords: Hypoxia; Maize kernel; Optical sensor; Oxygen imaging; Planar oxygen sensor; Respiration mapping; Seed.

MeSH terms

  • Calibration
  • Cell Respiration
  • Imaging, Three-Dimensional
  • Microscopy / methods*
  • Oxygen / metabolism*
  • Oxygen Consumption
  • Time Factors
  • Zea mays / metabolism

Substances

  • Oxygen