Wavelength-dependent differential interference contrast microscopy: selectively imaging nanoparticle probes in live cells

Anal Chem. 2009 Nov 15;81(22):9203-8. doi: 10.1021/ac901623b.

Abstract

Gold and silver nanoparticles display extraordinarily large apparent refractive indices near their plasmon resonance (PR) wavelengths. These nanoparticles show good contrast in a narrow spectral band but are poorly resolved at other wavelengths in differential interference contrast (DIC) microscopy. The wavelength dependence of DIC contrast of gold/silver nanoparticles is interpreted in terms of Mie's theory and DIC working principles. We further exploit this wavelength dependence by modifying a DIC microscope to enable simultaneous imaging at two wavelengths. We demonstrate that gold/silver nanoparticles immobilized on the same glass slides through hybridization can be differentiated and imaged separately. High-contrast, video-rate images of living cells can be recorded both with and without illuminating the gold nanoparticle probes, providing definitive probe identification. Dual-wavelength DIC microscopy thus presents a new approach to the simultaneous detection of multiple probes of interest for high-speed live-cell imaging.

Publication types

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

MeSH terms

  • Gold / analysis*
  • HeLa Cells
  • Humans
  • Metal Nanoparticles / analysis*
  • Microscopy, Interference / methods*
  • Peptide Fragments / chemistry*
  • Silver / analysis*
  • tat Gene Products, Human Immunodeficiency Virus / chemistry*

Substances

  • Peptide Fragments
  • tat Gene Products, Human Immunodeficiency Virus
  • tat peptide (47-57), Human immunodeficiency virus 1
  • Silver
  • Gold