Photonic force microscope based on optical tweezers and two-photon excitation for biological applications

J Struct Biol. 1997 Jul;119(2):202-11. doi: 10.1006/jsbi.1997.3880.

Abstract

A new scanning probe microscope, the photonic force microscope (PFM), based on optical tweezers and two-photon absorption processes for biological applications is described. Optical tweezers are used to trap a fluorescent latex bead with a diameter of 200 nm in an aqueous solution in all three dimensions. The fluorescent dye is chosen to fulfill the two-photon absorption criterion for the 1064-nm line of a Nd:YVO4 laser. The intensity of the fluorescence emission is utilized as a very sensitive position sensor along the optical axis. Two-dimensional images are formed by laterally scanning the trapped latex bead across biological samples while recording the two-photon-induced fluorescences intensity. A scanning probe image of the outer surface of a small neurite from a cultured rat hippocampal neuron is shown, which is hardly visible under differential interference contrast microscopy. The lateral resolution is given by the bead diameter; the axial resolution is 40 nm. Under the experimental conditions the maximal imaging force applied by the probe is below 5 pN.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Fluorescence
  • Fluorescent Dyes
  • Hippocampus / embryology
  • Image Processing, Computer-Assisted
  • Lasers
  • Latex
  • Microscopy / instrumentation
  • Microscopy / methods*
  • Microspheres
  • Neurons / ultrastructure*
  • Particle Size
  • Photons
  • Rats

Substances

  • Fluorescent Dyes
  • Latex