Near-field penetrating optical microscopy: a live cell nanoscale refractive index measurement technique for quantification of internal macromolecular density

Opt Lett. 2012 Feb 15;37(4):506-8. doi: 10.1364/OL.37.000506.

Abstract

Quantification of intracellular nanoscale macromolecular density distribution is a fundamental aspect to understanding cellular processes. We report a near-field penetrating optical microscopy (NPOM) technique to directly probe the internal nanoscale macromolecular density of biological cells through quantification of intracellular refractive index (RI). NPOM inserts a tapered optical fiber probe to successive depths into an illuminated sample. A 50 nm diameter probe tip collects signal that exhibits a linear relationship with the sample RI at a spatial resolution of approximately 50 nm for biologically relevant measurements, one order of magnitude finer than the Abbe diffraction limit. Live and fixed cell data illustrate the mechanical ability of a 50 nm probe to penetrate biological samples.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cells / ultrastructure*
  • Cheek
  • Humans
  • Macromolecular Substances / ultrastructure
  • Microscopy / methods*
  • Microscopy, Scanning Probe / methods
  • Optical Fibers
  • Refractometry / methods

Substances

  • Macromolecular Substances