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Nonlinear microscopes have seen an increase in popularity in the life sciences due to their molecular and structural specificity, high resolution, large penetration depth, and volumetric imaging capability. Nonetheless, the inherently weak optical signals demand long exposure times for live cell imaging. Here, by modifying the optical layout and illumination parameters, we can follow the rotation and translation of noncentrosymetric crystalline particles, or nanodoublers, with 50 μs acquisition times in living cells. The rotational diffusion can be derived from variations in the second harmonic intensity that originates from the rotation of the nanodoubler crystal axis. We envisage that by capitalizing on the biocompatibility, functionalizability, stability, and nondestructive optical response of the nanodoublers, novel insights on cellular dynamics are within reach.

Probing Rotational and Translational Diffusion of Nanodoublers in Living Cells on Microsecond Time Scales