The pulses that exit the interferometer were tailored to satisfy the optimum conditions to maximize the CARS signal intensity. To optimize the temporal pulse condition, the pulses were prechirped with a pair of dispersion-compensation mirrors (DCMP175, Thorlabs) so that they were compressed down to near-transform–limited pulses at cells in the microfluidic channel. In our system, the pulses were compressed down to 18 fs at cells in the microfluidic channel by adding a negative second-order dispersion of 2450 fs2 to the pulses. To optimize the spectral pulse condition, the pulses were filtered by a long-pass optical filter with a cutoff wavelength of 750 nm (FELH0750, Thorlabs). Suppressing the high-frequency edge of the spectrum of the pump and probe pulses ensures noise reduction for the CARS signal detection.

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