Compact simultaneous label-free autofluorescence multi-harmonic microscopy for user-friendly photodamage-monitored imaging.

Publisher: Published by SPIE--the International Society for Optical Engineering in cooperation with International Biomedical Optics Society Country of Publication: United States NLM ID: 9605853 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1560-2281 (Electronic) Linking ISSN: 10833668 NLM ISO Abbreviation: J Biomed Opt Subsets: MEDLINE

Original Publication: Bellingham, WA : Published by SPIE--the International Society for Optical Engineering in cooperation with International Biomedical Optics Society, c1996-

Lasers* ; Photons*; Nonlinear Optical Microscopy ; Microscopy, Fluorescence, Multiphoton/methods

Significance: Label-free nonlinear optical microscopy has become a powerful tool for biomedical research. However, the possible photodamage risk hinders further clinical applications.
Aim: To reduce these adverse effects, we constructed a new platform of simultaneous label-free autofluorescence multi-harmonic (SLAM) microscopy, featuring four-channel multimodal imaging, inline photodamage monitoring, and pulse repetition-rate tuning.
Approach: Using a large-core birefringent photonic crystal fiber for spectral broadening and a prism compressor for pulse pre-chirping, this system allows users to independently adjust pulse width, repetition rate, and energy, which is useful for optimizing imaging conditions towards no/minimal photodamage.
Results: It demonstrates label-free multichannel imaging at one excitation pulse per image pixel and thus paves the way for improving the imaging speed by a faster optical scanner with a low risk of nonlinear photodamage. Moreover, the system grants users the flexibility to autonomously fine-tune repetition rate, pulse width, and average power, free from interference, ensuring the discovery of optimal imaging conditions with high SNR and minimal phototoxicity across various applications.
Conclusions: The combination of a stable laser source, independently tunable ultrashort pulse, photodamage monitoring features, and a compact design makes this new system a robust, powerful, and user-friendly imaging platform.
(© 2024 The Authors.)

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P41 EB031772 United States EB NIBIB NIH HHS; R01 CA241618 United States CA NCI NIH HHS

Keywords: label-free imaging; multiphoton microscopy; nonlinear fiber optics; photodamage; tunable ultrashort pulse

Date Created: 20240315 Date Completed: 20240318 Latest Revision: 20241104

20241104

PMC10939229

10.1117/1.JBO.29.3.036501

38487259