For deep imaging depth and least invasiveness, people commonly use 1100-1300 nm femtosecond laser sources to perform label-free in vivo microscopy. The modalities include reflectance confocal, two & three photon fluorescence, and second & third harmonic generation microscopy. However, most of the laser sources are typically based on bulky oscillators, which are sensitive to environment conditions and less stable for routine clinical use. In contrast, fiber-based lasers have simpler cavity design and potentially compact size for movable use. In this presentation, we demonstrate a fiber-based 1150 nm femtosecond laser source, with 6.5 nJ pulse energy, 86 fs pulse-width, and 11.25 MHz pulse repetition rate. It was achieved by a Bismuth Borate (BIBO) or Magnesium-doped periodically poled Lithium Niobate (MgO:PPLN) mediated frequency doubling of the 2300 nm solitons, generated from an excitation of 1550 nm Er:fiber femtosecond laser pulses on a large mode area photonic crystal fiber. Combined with a laser scanned microscope and a home-build data acquisition card, we achieve a pulse-per-pixel harmonic generation microscopy in vivo at a 30 Hz frame rate. In the future, this solution is potential to be used for label-free clinical virtual optical biopsy.
Harmonic generation microscopy (HGM) has become one unique tool of optical virtual biopsy for the diagnosis of cancer and the in vivo cytometry of leukocytes. Without labeling, HGM can reveal the submicron features of tissues and cells in vivo. For deep imaging depth and minimal invasiveness, people commonly adopt 1100- to 1300-nm femtosecond laser sources. However, those lasers are typically based on bulky oscillators whose performances are sensitive to environmental conditions. We demonstrate a fiber-based 1150-nm femtosecond laser source, with 6.5-nJ pulse energy, 86-fs pulse width, and 11.25-MHz pulse repetition rate. It was obtained by a bismuth borate or magnesium-doped periodically poled lithium niobate (MgO:PPLN) mediated frequency doubling of the 2300-nm solitons, generated from an excitation of 1550-nm femtosecond pulses on a large mode area photonic crystal fiber. Combined with a home-built laser scanned microscope and a tailor-made frame grabber, we achieve a pulse-per-pixel HGM imaging in vivo at a 30-Hz frame rate. This integrated solution has the potential to be developed as a stable HGM system for routine clinical use.
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