Physical chemistry in a single live cell: confocal microscopy

Phys Chem Chem Phys. 2017 May 24;19(20):12620-12627. doi: 10.1039/c7cp02228j.

Abstract

A live cell is a complex, yet extremely important container. Understanding the dynamics in a selected intracellular component is a challenging task. We have recently made significant progress in this direction using a confocal microscope as a tool. The smallest size of the focused spot in a confocal microscope is ∼0.2 μm (200 nm). This is nearly one hundred times smaller than the size of a live cell. Thus, one can selectively study different intracellular components/organelles in a live cell. In this paper, we discuss how one can image different intracellular components/organelles, record fluorescence spectra and decay at different locations, ascertain local polarity and viscosity, and monitor the dynamics of solvation, proton transfer, red-ox and other phenomena at specified locations/organelles inside a cell. We will highlight how this knowledge enriched us in differentiating between cancer and non-cancer cells, 3D tumor spheroids and towards drug delivery.

MeSH terms

  • Amyloid beta-Peptides / chemistry
  • Cell Line
  • Coumarins / chemistry
  • Fluorescent Dyes / chemistry
  • Humans
  • Microscopy, Confocal*
  • Organelles / chemistry*
  • Organelles / pathology
  • Protons
  • Spheroids, Cellular / chemistry
  • Spheroids, Cellular / pathology
  • Viscosity

Substances

  • Amyloid beta-Peptides
  • Coumarins
  • Fluorescent Dyes
  • Protons
  • N-(4-(7-diethylamino-4-methylcoumarin-3-yl)phenyl)maleimide