Nonlinear Dark-Field Microscopy

Abstract

Dark-field microscopy is a background-free imaging method that provides high sensitivity and a large signal-to-noise ratio. It finds application in nanoscale detection, biophysics and biosensing, particle tracking, single molecule spectroscopy, X-ray imaging, and failure analysis of materials. In dark-field microscopy, the unscattered light path is typically excluded from the angular range of signal detection. This restriction reduces the numerical aperture and affects the resolution. Here we introduce a nonlinear dark-field scheme that overcomes this restriction. Two laser beams of frequencies ω1 and ω2 are used to illuminate a sample surface and to generate a purely evanescent field at the four-wave mixing (4WM) frequency ω4wm = 2ω1 − ω2. The evanescent 4WM field scatters at sample features and generates radiation that is detected by standard far-field optics. This nonlinear dark-field scheme works with samples of any material and is compatible with applications ranging from biological imaging to failure analysis.© 2010 American Chemical Society.