Cell Volume Measurements by Optical Transmission Microscopy

Curr Protoc Cytom. 2019 Sep;90(1):e62. doi: 10.1002/cpcy.62.

Abstract

Cell volume is an important parameter in studying cell adaptation to anisosmotic stress, activation of monovalent ion channels, and cell death. This article describes a method for measurement of the volumes of adherent cells using a standard light microscope. A coverslip with attached cells is placed in a shallow chamber in a medium containing a strongly absorbing and cell-impermeant dye, Acid Blue 9. When such a sample is imaged in transmitted light at a wavelength of maximum dye absorption (630 nm), the resulting contrast quantitatively reflects cell thickness; once the thickness is known at every point, the volume can be computed as well. Technical details, interpretation of data, and possible artifacts are discussed. © 2019 by John Wiley & Sons, Inc.

Keywords: Acid Blue 9; cell volume; optical microscopy; transmission-through-dye microscopy.

MeSH terms

  • Benzenesulfonates / chemistry*
  • Cell Line
  • Cell Size*
  • Humans
  • Image Processing, Computer-Assisted*
  • Microscopy*
  • Staining and Labeling*

Substances

  • Benzenesulfonates
  • brilliant blue