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Label-free nonlinear optical microscopy has become a powerful tool forbiomedical research. However, the possible photodamage risk hinder furtherclinical applications. To reduce these adverse effects, we constructed a newplatform of simultaneous label-free autofluorescence multiharmonic (SLAM)microscopy, featuring 5-channel multimodal imaging, inline photodamagemonitoring, and pulse repetition-rate tuning. By the use of a birefringentphotonic crystal fiber for spectral broadening (rather than supercontinuumgeneration) and a prism compressor for pulse pre-chirping, this system allowsusers to independently adjust pulse width, repetition rate, and energy, whichis useful for optimizing imaging condition towards no/minimal photodamage.Also, it demonstrates label-free multichannel imaging at one excitation pulseper image pixel and thus paves the way for improving the imaging speed by afaster optical scanner.

Compact simultaneous label-free autofluorescence multi-harmonic (SLAM) microscopy for user-friendly photodamage-monitored imaging