Axial resolution enhancement of third harmonic generation microscopy by harmonic focal point axial modulation

Abstract

Harmonic focal point axial modulation (h-FPAM) in nonlinear optical microscopy is introduced and used to enhance the axial resolution and the signal to background ratio by modulating the focal point of a beam of femtosecond pulses train along the axial direction and phase sensitively filtering the resulting signals using a lock-in amplifier. Axial resolution enhancement factor of 2.05 is acquired in 2f detection mode for the third harmonic generation microscopy of glass-oil interfaces. This technique also resolves the image interpretation problem of the fundamental harmonic FPAM technique. Moreover, the potential of this technique for axial sectioning is demonstrated by acquiring images from a red blood cell.