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Photoacoustics
2020 Jun 01;18:100163. doi: 10.1016/j.pacs.2020.100163.
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All-fibre supercontinuum laser for in vivo multispectral photoacoustic microscopy of lipids in the extended near-infrared region.
Dasa MK
,
Nteroli G
,
Bowen P
,
Messa G
,
Feng Y
,
Petersen CR
,
Koutsikou S
,
Bondu M
,
Moselund PM
,
Podoleanu A
,
Bradu A
,
Markos C
,
Bang O
.
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Among the numerous endogenous biological molecules, information on lipids is highly coveted for understanding both aspects of developmental biology and research in fatal chronic diseases. Due to the pronounced absorption features of lipids in the extended near-infrared region (1650-1850 nm), visualisation and identification of lipids become possible using multi-spectral photoacoustic (optoacoustic) microscopy. However, the spectroscopic studies in this spectral region require lasers that can produce high pulse energies over a broad spectral bandwidth to efficiently excite strong photoacoustic signals. The most well-known laser sources capable of satisfying the multi-spectral photoacoustic microscopy requirements (tunability and pulse energy) are tunable nanosecond optical parametric oscillators. However, these lasers have an inherently large footprint, thus preventing their use in compact microscopy systems. Besides, they exhibit low-repetition rates. Here, we demonstrate a compact all-fibre, high pulse energy supercontinuum laser that covers a spectral range from 1440 to 1870 nm with a 7 ns pulse duration and total energy of 18.3 μJ at a repetition rate of 100 kHz. Using the developed high-pulse energy source, we perform multi-spectral photoacoustic microscopy imaging of lipids, both ex vivo on adipose tissue and in vivo to study the development of Xenopus laevis tadpoles, using six different excitation bands over the first overtone transition of C-H vibration bonds (1650-1850 nm).
Fig. 1. (a) Schematic of the MS-PAM system. C1: reflective collimator, LVF: linear variable filter, GM: galvo-mirrors, L1: achromatic lens, MS: microscopy slide, FT: flat transducer, PC: the personal computer. The photograph in the bottom left shows the all-fibre SC laser and the power supply unit with a scale bar of 10 cm. (b): Lateral resolution of the MS-PAM system estimated by using the edge and line spread functions. (c): PAM image of the USAF resolution target at 1720 nm.
Fig. 2. (a) Schematic of the SC laser configuration. DCF4: dispersion-shifted fibre. (b): PSD of preamplifier, booster amplifier and the SC spectrum generated by pumping a 4.5 m non-zero DSF. (c): SC output spectrum (solid line) in comparison with the SC reported in our previous work (dashed line). (d): SC Excitation bands filtered using the LVF. (e): Pulse energy and bandwidth of filtered excitation bands from the SC laser.
Fig. 3. Optical image and MS-PAM images of ex vivo adipose tissue. Six z-projected en-face MS-PAM images are acquired from 1600 nm to 1800 nm in steps of 40 nm. The white bar in the optical image and the last MS-PAM image at 1800 nm represents the scale bar of 1 mm.
Fig. 4. (a) Normalized PA amplitudes at two different regions (labelled 1 and 2 in the inset) for the six excitation bands with corresponding PA amplitudes for adipose tissue measurements reported in our previous study [33]. The area of each region is 0.087 mm2. (b) Measured spectral variation of the SNR of ex vivo adipose tissue measured in MS-PAM images.
Fig. 5. Optical image and 6 in vivo z-projected en-face MS-PAM images of a Xenopus laevis tadpole acquired from 1600 nm to 1800 nm in steps of 40 nm. The highlighted region in the optical image shows the yolk sac. The scale bar represents 1 mm.
Fig. 6. (a) Normalized PA amplitudes of a small region (about 0.087 mm2) in the MS-PAM images of a tadpole at three different places (labelled 1, 2, and 3) for the six excitation bands. (b) Measured SNR of tadpole MS-PAM images at all the six excitation bands.
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