Multispectral Photoacoustic Microscopy Research Articles

Electronically Controlled Dual‐Wavelength Switchable SRS Fiber Amplifier in the NIR‐II Region for Multispectral Photoacoustic Microscopy

AbstractPhotoacoustic microscopy (PAM) is a high‐resolution and non‐invasive imaging modality that provides optical absorption contrast. By employing dual‐ or multiple‐wavelength excitation, PAM extends its capabilities to offer valuable spectroscopic information. To achieve efficient multispectral PAM imaging, an essential requirement is a light source characterized by a high repetition rate and switching rate, a ≈microjoule pulse energy, and a ≈nanosecond pulse duration. However, there exists a notable deficiency in suitable light sources, particularly in the near‐infrared‐II window. In this study, a custom‐built all‐fiber‐based light source is reported that provides >3 µJ, 2 ns pulses with a repetition rate of 200 kHz. Digitally addressed semiconductor seed lasers, followed by stimulated Raman scattering amplification, enabled arbitrary sequences of pulses having wavelengths of either 1168.4 or 1202.1 nm. In a switching mode of operation, a 100 kHz switching rate is used to alternate between these wavelengths in even/odd pulses. Furthermore, a high‐resolution multispectral photoacoustic microscopy of three polymer samples is demonstrated with the proposed light source.

Dual-wavelength UV-visible metalens for multispectral photoacoustic microscopy: A simulation study

Photoacoustic microscopy is advancing with research on utilizing ultraviolet and visible light. Dual-wavelength approaches are sought for observing DNA/RNA- and vascular-related disorders. However, the availability of high numerical aperture lenses covering both ultraviolet and visible wavelengths is severely limited due to challenges such as chromatic aberration in the optics. Herein, we present a groundbreaking proposal as a pioneering simulation study for incorporating multilayer metalenses into ultraviolet-visible photoacoustic microscopy. The proposed metalens has a thickness of 1.4 µm and high numerical aperture of 0.8. By arranging cylindrical hafnium oxide nanopillars, we design an achromatic transmissive lens for 266 and 532 nm wavelengths. The metalens achieves a diffraction-limited focal spot, surpassing commercially available objective lenses. Through three-dimensional photoacoustic simulation, we demonstrate high-resolution imaging with superior endogenous contrast of targets with ultraviolet and visible optical absorption bands. This metalens will open new possibilities for downsized multispectral photoacoustic microscopy in clinical and preclinical applications.

Quickly Alternating Green and Red Laser Source for Real-time Multispectral Photoacoustic Microscopy

Multispectral photoacoustic microscopy uses a wavelength-dependent absorption difference as a contrast mechanism to image the target molecule. In this paper, we present a novel multispectral pulsed fiber laser source, which selectively alternates the excitation wavelengths between green and red colors based on the stimulated Raman scattering (SRS) effect for imaging. This laser has a high pulse repetition rate (PRR) of 300 kHz and high pulse energy of more than 200 nJ meeting the real-time requirements of optical-resolution photoacoustic microscopy imaging. By switching the polarization state of the pump light and optical paths of the pump light, the operating wavelengths of the light source can be selectively alternated at the same fast PRR for any two SRS peak wavelengths between 545 and 655 nm. At 545 nm excitation wavelength, molecular photoacoustic signals from both blood vessels and gold nanorods were obtained simultaneously. However, at 655 nm, the photoacoustic signals of gold nanorods were dominant because the absorption of light by the blood vessels decreased drastically in the spectral region over 600 nm. Thus the multispectral photoacoustic system designed using the novel laser source implemented here could simultaneously monitor the time-dependent fast movement of two molecules independently, having different wavelength-dependent absorption properties at a high repetition rate of 0.49 frames per second (fps).

All-fibre supercontinuum laser for in vivo multispectral photoacoustic microscopy of lipids in the extended near-infrared region

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).

Co-impulse multispectral photoacoustic microscopy and optical coherence tomography system using a single supercontinuum laser.

A combination of multispectral photoacoustic microscopy (PAM) and optical coherence tomography (OCT) by a single light source was previously realized discretely; however, this is unfavorable for visualizing vital physiological and pathological activities in vivo. Here, a co-impulse dual-mode imaging system that simultaneously enables multispectral PAM and OCT using a megahertz supercontinuum pulse laser in vivo is presented. The 500-600nm band is used for functional PAM imaging, which can flexibly switch between different wavelengths, while the 600-840nm band is selected for OCT imaging. A mimicking phantom experiment and in vivo imaging of normal and melanoma mouse ears demonstrate that the co-impulse multispectral PAM-OCT system can simultaneously provide structural and functional information for bioimaging.

Multispectral photoacoustic microscopy and optical coherence tomography using a single supercontinuum source

We report on the use of a single supercontinuum (SC) source for multimodal imaging. The 2-octave bandwidth (475–2300nm) makes the SC source suitable for optical coherence tomography (OCT) as well as for multispectral photoacoustic microscopy (MPAM). The IR band centered at 1310nm is chosen for OCT to penetrate deeper into tissue with 8mW average power on the sample. The 500–840nm band is used for MPAM. The source has the ability to select the central wavelength as well as the spectral bandwidth. An energy of more than 35nJ within a less than 50nm bandwidth is achieved on the sample for wavelengths longer than 500nm. In the present paper, we demonstrate the capabilities of such a multimodality imaging instrument based on a single optical source. In vitro mouse ear B-scan images are presented.

Near-infrared multispectral photoacoustic microscopy using a graded-index fiber amplifier

We demonstrate optical resolution photoacoustic microscopy (OR-PAM) of lipid-rich tissue using a multi-wavelength pulsed laser based on nonlinear fiber optics. 1047nm laser pulses are converted to 1098, 1153, 1215, and 1270nm pulses via stimulated Raman scattering in a graded-index multimode fiber. Multispectral PAM of a lipid phantom is demonstrated with our low-cost and simple technique.

Single all-fiber-based nanosecond-pulsed supercontinuum source for multispectral photoacoustic microscopy and optical coherence tomography

We report the usefulness of a single all-fiber-based supercontinuum (SC) source for combined photoacoustic microscopy (PAM) and optical coherence tomography (OCT). The SC light is generated by a tapered photonic crystal fiber pumped by a nanosecond pulsed master oscillator power amplifier at 1064 nm. The spectrum is split into a shorter wavelength band (500-800 nm) for single/multi-spectral PAM and a longer wavelength band (800-900 nm) band for OCT. In vivo mouse ear imaging was achieved with an integrated dual-modality system. We further demonstrated its potential for spectroscopic photoacoustic imaging by doing multispectral measurements on retinal pigment epithelium and blood samples with 15-nm linewidth.

High energy supercontinuum sources using tapered photonic crystal fibers for multispectral photoacoustic microscopy.

We demonstrate a record bandwidth high energy supercontinuum source suitable for multispectral photoacoustic microscopy. The source has more than 150 nJ/10 nm bandwidth over a spectral range of 500 to 1600 nm. This performance is achieved using a carefully designed fiber taper with large-core input for improved power handling and small-core output that provides the desired spectral range of the supercontinuum source.