Heterodyne detected nonlinear optical imaging in a lock-in free manner

We report a compact, cost-effective tuned amplifier for frequency-selective amplification of the modulated signal in heterodyne detected nonlinear optical microscopy. Our method improved the signal to noise ratio by an order of magnitude compared to conventional lock-in detection, as demonstrated through stimulated Raman scattering imaging of live cells and tissues at the speed of 2 μsec/pixel. Application of the tuned amplifier to transient absorption microscopy is also demonstrated. The increased signal to noise ratio allowed epi-detected in vivo imaging of myelin and blood in rat spinal cord with high spatial resolution.