Phase-resolved wide-field CARS microscopy with speckle illumination

Coherent anti-Stokes Raman scattering (CARS) microscopy enables label-free chemical imaging of biological samples and materials. Conventionally, CARS is implemented using a point-scanning approach that probes a single vibrational mode at a time. Hyperspectral CARS enhances chemical specificity by sequentially addressing multiple Raman modes. However, the measured CARS intensity is distorted by an undesired non-resonant background, which broadens and shifts the Raman peaks, thereby hindering the interpretability of hyperspectral images. Here, we introduce a phase-sensitive, wide-field hyperspectral CARS microscopy scheme that suppresses the non-resonant background. The proposed approach combines three key components: a high-power picosecond tunable optical parametric amplifier (OPA), speckle illumination, and quantitative phase imaging based on quadriwave lateral shearing interferometry (QLSI). The high-power OPA provides the peak power required for efficient nonlinear excitation. Speckle illumination distributes the optical energy over the objective back pupil and mitigates coherent imaging artifacts. QLSI enables the simultaneous measurement of the amplitude and phase of the CARS field, thereby allowing separation of resonant and non-resonant contributions, without the need for an external reference beam. This unique combination enables practical phase-resolved CARS imaging over a field of view exceeding $60\times 60{\text{textmu}\text{m}}^2$ at a frame rate of 1.4 Hz. We illustrate the approach by acquiring hyperspectral images of microplastics and liver steatosis across the entire CH-stretching region.