The Fabry-Perot (FP) sensor is typically read out by scanning a focused laser beam over the sensor surface and measuring the reflected light. However, the acquisition time can be relatively long when a large number of spatial sampling points are required. An alternative approach is to use widefield illumination and measure the reflected light using a camera. This approach has enabled the acquisition of 3D photoacoustic images composed of 327,000 spatial sampling points in a couple of seconds or less. In this study, we investigate the possibility of further improving imaging speed by combining the widefield illumination approach with the principles of compressed sensing. Preliminary results suggest that current imaging times could be reduced by at least a factor of 4, enabling the acquisition of 3D photoacoustic images in sub-second timeframes.
KEYWORDS: 3D scanning, 3D image processing, Laser scanners, Stereoscopy, Photoacoustic spectroscopy, Imaging systems, 3D acquisition, Visualization, Optical scanning systems, Video
A 3D high resolution Fabry Perot photoacoustic scanner has been developed for clinical use. The system now employs a novel 64-channel optical scanning architecture and compressed sensing methods providing up to three orders of magnitude faster acquisition than previous pre-clinical embodiments. To demonstrate the rapid, high quality volumetric imaging capabilities, and the versatility of the scanner, images from different regions of the body that are known to exhibit distinctive vascular anatomy were acquired using healthy volunteers as participants. In addition, to illustrate potential clinical applicability, imaging studies of patients suffering from diseases characterised by abnormal vascular anatomy were conducted.
Fabry-Perot scanners are typically read-out in a sequential manner. An alternative approach is to use a widefield illumination and measure the reflected light in parallel using a camera. This approach allows for a finer spatial sampling of the Fabry-Perot sensor and can speed up the image acquisition. In a first instance, the capability of the camera-based approach to finely spatially sample the sensor, with the means of improving image resolution, was demonstrated. In a second instance, the ability of the camera-based system to acquire 3D photoacoustic images in 200 ms when operating at a PRF of 1 kHz was demonstrated.
Photoacoustic (PA) wavefront shaping (WS; PAWS) could allow focusing light deep in biological tissue. This could enable increasing the penetration depth of biomedical optical techniques including PA imaging. However, focussing at depth requires a light source of long coherence length (CL), presenting a challenge because the CLs of typical PA excitation lasers are short. To address this challenge, we developed a PAWS system based on an externally modulated external cavity laser with a long CL. The system was demonstrated by focussing light through rigid scattering media using both PAWS and optical WS. PAWS enabled focussing through diffusers with 8 × enhancements, while all-optical WS enabled focussing through various scattering media including a 5.8 mm thick tissue phantom. By enabling PAWS with increased coherence, the system could facilitate exploring the practical depth limits of PAWS, paving the way to focussing light deep in tissue.
Fabry-Perot (FP) based scanners are typically interrogated by scanning a single focused laser beam over its surface and measuring the reflected light. However, this approach can be relatively slow with acquisition times on the order of minutes. An alternative is to parallelise the read-out of the Fabry-Perot sensor by illuminating it with a large diameter collimated interrogation beam (e.g. 1 cm) and measuring the reflected light using an InGaAs camera. This approach allows acquiring 3D photoacoustic images in sub-second time frames. To demonstrate this speed advantage, a camera based FP scanner was developed and evaluated.
A 3D high resolution scanner has been developed specifically for clinical use. The novel scanner architecture employing multiple interrogation beams can acquire a 3D image in less than 1 second. An initial technical validation study has been undertaken in human volunteers to determine repeatability, reproducibility and patient acceptability. Thereafter, a first-in-man clinical study aimed at assessing diagnostic accuracy in patients with inflammatory diseases has been completed.
The imaging speed of optical resolution photoacoustic microscopy (OR-PAM) using pulsed excitation is fundamentally limited by the range ambiguity condition, which defines the maximum laser pulse repetition frequency (PRF). To operate at this theoretical upper limit and maximize acquisition speed, a custom-built fiber laser capable of operating at a PRF of up to 2 MHz was combined with a fast laser scanning optical OR-PAM system based on a stationary fiber-optic ultrasound sensor. A large area (10 mm × 10 mm) of the mouse ear was imaged within 8 s, when acquiring 16 million A-lines and operating the laser at a PRF of 2 MHz. This corresponds to a factor of four improvement in imaging speed compared to the fastest OR-PAM system previously reported. The ability to operate at high-imaging frame rates also allows the capture of hemodynamic events such as blood flow. It is considered that this system offers opportunities for high throughput imaging and visualizing dynamic physiological events using OR-PAM.
Photoacoustic signals are typically generated using Q-switched Nd:YAG pumped OPO systems, as they can provide the necessary nanosecond pulse durations with mJ pulse energies required for photoacoustic tomography. However, these sources are often bulky, require external water cooling and regular maintenance and provide low pulse repetition frequencies (PRF<100Hz) thus limiting image frame rate.
Fibre lasers can overcome these limitations and additionally offer much greater flexibility in their temporal output characteristics (e.g. pulse shaping and duration). Although fibre lasers have been used in optical-resolution photoacoustic microscopy, they have found limited application in widefield photoacoustic tomography (PAT) due to the relatively low pulse energy (<1mJ) provided by commercial systems. These low pulse energies are a consequence of small core diameter (<25m) fibres required to achieve a high beam quality. However, for widefield PAT, high beam quality is not a requirement and therefore fibre lasers with larger core diameters (>100m) can be used, enabling significantly higher pulse energies (>10mJ) to be achieved.
A novel compact fibre laser which uses a custom drawn large core diameter fibre (100m) to provide high pulse energies (15mJ) and variable PRFs (100Hz-1kHz) and pulse durations (10-400ns) has been developed and evaluated. The fibre laser was combined with a fast Fabry Perot (FP) scanner in order to evaluate its suitability for PAT of biological tissue. The high PRF (>400Hz) of the laser has allowed tomographic images of the microvasculature of the palm of a hand to be obtained in less than one second, significantly quicker than previously achieved with a FP scanner. In addition, the ability to arbitrarily vary the temporal shape of the laser pulse offers new opportunities for controlling the acoustic frequency content of the photoacoustic signal in order to optimise penetration depth and image resolution. For example, the laser pulse duration can be increased in order to shift the acoustic frequency components to lower frequencies which are less attenuated by tissue acoustic absorption and thus improve SNR. To investigate these concepts, a tissue mimicking phantom was imaged for a range of tailored excitation pulses (e.g. different pulse durations, trains of pulses) and their effect on the contrast to noise ratio (CNR) and image resolution observed.
A novel compact fibre laser, able to provide higher pulse energies (>10mJ) than previously reported and with enhanced functionality is presented. It is demonstrated that fibre lasers are a viable alternative to standard Q-switched lasers for photoacoustic tomographic applications in medicine and biology.
Two custom fibre lasers have been developed. One is designed for widefield photoacoustic tomography (PAT) and uses a custom drawn large core diameter fibre (100μm) to provide high pulse energies (5mJ). It also provides a variable pulse repetition frequency (100Hz-400Hz) and pulse duration (10-150ns) and is compact (of comparable dimensions to a desktop PC) and does not require external water cooling. This system was used to acquire in vivo images of the subcutaneous microvasculature in the human palm. The second laser is designed for Optical Resolution Photoacoustic Microscopy (OR-PAM) and provides a high quality beam (M2<1.1), pulse energies >1μJ with a pulse repetition frequency (PRF) up to 2MHz, and a 532nm emission wavelength. The high PRF of this laser was exploited for ultra-fast image acquisition. The compact size and enhanced functionality of these lasers offers a major opportunity to facilitate the translation of photoacoustic imaging to practical applications in medicine and biology.
KEYWORDS: Lymphatic system, Photoacoustic imaging, Cancer, Photoacoustic spectroscopy, Absorption, Visualization, Sensors, 3D image processing, Magnetic resonance imaging, Signal to noise ratio
Lymph nodes play a central role in metastatic cancer spread and are a key clinical assessment target. Abnormal node vascularization, morphology, and size may be indicative of disease but can be difficult to visualize with sufficient accuracy using existing clinical imaging modalities. To explore the potential utility of photoacoustic imaging for the assessment of lymph nodes, images of ex vivo samples were obtained at multiple wavelengths using a high-resolution three-dimensional photoacoustic scanner. These images showed that hemoglobin based contrast reveals nodal vasculature and lipid-based contrast reveals the exterior node size, shape, and boundary integrity. These two sources of complementary contrast may allow indirect observation of cancer, suggesting a future role for photoacoustic imaging as a tool for the clinical assessment of lymph nodes.
KEYWORDS: Sensors, Photoacoustic microscopy, Fiber optics sensors, Ultrasonography, Fiber optics, Photoacoustic spectroscopy, Signal detection, Acoustics, Absorption, Signal to noise ratio
Laser-Scanning-Optical-Resolution Photoacoustic Microscopy (LSOR-PAM) requires an ultrasound detector with a low noise equivalent pressure (NEP) and a large angular detection aperture in order to image a large field of view (FOV). It is however challenging to meet these requirements when using piezoelectric receivers since using a small sensing element size (<100μm) in order to achieve a large angular detection aperture will inevitability reduce the sensitivity of the detector as it scales with decreasing element size. Fibre optic ultrasound sensors based on a Fabry Perot cavity do not suffer from this limitation and can provide high detection sensitivity (NEP<0.1kPa over a 20 MHz measurement bandwidth) with a large angular detection aperture due to their small active element size (~10μm). A LSOR-PAM system was developed and combined with this type of fibre optic ultrasound sensor. A set of phantom studies were undertaken. The first study demonstrated that a high resolution image over a large field of view (Ø11mm) could be obtained with a sampledetector separation of only 1.6mm. In the second study, a 12μm diameter tube filled with methylene blue whose absorption coefficient was similar to that of blood was visualised demonstrating that the fibre optic sensor could provide sufficient SNR for in-vivo microvascular OR-PAM imaging. These preliminary results suggest that the fibre optic sensor has the potential to outperform piezoelectric detectors for Laser-Scanning Optical Resolution Photoacoustic Microscopy (LSOR-PAM).
A simple method of characterizing organic dyes and nanoparticles used as contrast agents for photoacoustic molecular imaging based on relative photoacoustic measurements is described. By acquiring just two time-resolved photoacoustic signals, one in the sample of interest and the other in water, measurements of the thermalisation efficiency and other parameters relevant to the characterization of contrast agents can be acquired. The method was validated using absorbing solutions of known thermalisation efficiency and Grüneisen coefficient. It was then used to measure the thermalisation efficiency of solutions of gold nanorods, rhodamine B, methylene blue, IR-820, fluorescein and cresyl violet. In addition, photoacoustic measurements of the photostability of these substances were acquired.
Semiconductor light sources, such as laser diodes or light emitting diodes (LEDs) could provide an inexpensive
and compact alternative to traditional Q-switched lasers for photoacoustic imaging. So far, only laser diodes 1-3
operating in the 750 to 905nm wavelength range have been investigated for this purpose. However, operating in
the visible wavelength range (400nm to 650nm) where blood is strongly absorbent (<10cm-1) and water
absorption is weak (<0.01cm-1) could allow for high contrast photoacoustic images of the superficial vasculature to be achieved. High power laser diodes (<10Watt peak power) are however not available in this wavelength
range. High power LEDs could be a potential alternative as they are widely available in the visible wavelength
range (400nm to 632nm) and relatively cheap. High power LEDs are generally operated in continuous wave
mode and provide average powers of several Watts. The possibility of over driving them by tens of times their
rated current when driven at a low duty cycle (<1%), offers the prospect of achieving similar pulse energies
(tens of μJ) to that provided by high peak power pulsed laser diodes. To demonstrate the possibility of using
high power LEDs as an excitation source for biomedical applications, single point measurements were
implemented in a realistic blood vessel phantom. A four colour device was also used to demonstrate the
possibility of using LEDs for making spectroscopic measurements. It was shown that when driving all four
wavelengths at once, the generated photoacoustic signal could be used to design a filter in order to improve the
SNR of the photoacoustic signals generated at each individual wavelength. The possibility of acquiring
multiwavelength data sets simultaneously when using Golay excitation methods was also demonstrated. This
preliminary study demonstrated the potential for using high power LEDs as an inexpensive and compact
excitation source for biomedical photoacoustics.
Spectroscopic photoacoustic imaging has the potential to discriminate between normal and lipid-rich atheromatous areas of arterial tissue by exploiting the differences in the absorption spectra of lipids and normal arterial tissue in the 740 to 1400 nm wavelength range. Identification of regions of high lipid concentration would be useful to identify plaques that are likely to rupture (vulnerable plaques). To demonstrate the feasibility of visualizing lipid-rich plaques, samples of human aortas were imaged in forward mode, at wavelengths of 970 and 1210 nm. It was shown that the structure of the arterial wall and the boundaries of lipid-rich plaques obtained from the photoacoustic images were in good agreement with histology. The presence of lipids was also confirmed by comparing the photoacoustic spectra (740 to 1400 nm) obtained in a region within the plaque to the spectral signature of lipids. Furthermore, a lipid-rich plaque was successfully imaged while illuminating the sample through 2.8 mm of blood demonstrating the possibility of implementing the photoacoustic technique in vivo.
KEYWORDS: Sensors, Acoustics, Photoacoustic spectroscopy, Signal to noise ratio, Signal attenuation, Tissue optics, Chemical elements, Signal detection, Photoacoustic imaging, Breast
For deep tissue photoacoustic imaging, piezoelectric ultrasound detectors with large element sizes (>1mm) and relatively
low centre frequencies (<5MHz) are generally used, as they can provide the required high sensitivity to achieve imaging
depths of several centimetres. However, these detectors are generally not optimised in terms of element size and
bandwidth. To identify these parameters in order to improve SNR and spatial resolution, two models were employed.
The first was a numerical model and was used to investigate the effect of varying the detector element size on the
amplitude and SNR of photoacoustic images. The second model was used to optimise the detector bandwidth. For this,
the frequency content of simulated photoacoustic signals were studied for a range of depths and acoustic source sizes.
The model was based on an analytical solution to the wave equation for a cylindrical source and incorporated the effects
of frequency dependent acoustic attenuation. These models provide a new framework for optimising the design of
photoacoustic scanners for breast and other deep tissue imaging applications.
The use of a pulsed fibre laser as an excitation source for photoacoustic tomography has been investigated. Fibre lasers
have the advantage of being compact, robust and efficient compared to traditional excitation sources used for
photoacoustic tomography (e.g. Q-switched Nd:YAG pumped OPO or dye systems). Their high pulse repetition
frequencies and adjustable pulse duration, shape and duty cycle also enables a wide range of time and frequency domain
excitation methods to be investigated. A 1060nm, 20W fibre laser was used to generate acoustic waves in a tissue
mimicking phantom composed of blood filled tubes immersed in a 1% solution of intralipid (μ's=1mm-1) . The laser was
then combined with a Fabry Perot photoacoustic imaging system to obtain 3D images of a tissue mimicking phantom and
an in vivo image of the vasculature of the palm of a volunteer. This study has demonstrated that pulsed fibre lasers have
potential application as an excitation source for photoacoustic imaging of superficial blood vessels.
Recently it has been shown that multiwavelength photoacoustic imaging has the potential to discriminate between
normal and atheromatous areas of arterial tissue when operating in the 740-1300nm wavelength range. At this
wavelength range the absorption spectrum of lipids and normal arterial tissue are significantly different allowing
discrimination between one another. Also, this wavelength range has the advantage of being relatively weakly
absorbed by blood. This obviates the need for a saline flush if implemented using an intravascular imaging
probe. In this study we investigate the possibility of identifying regions of high lipid concentration from 2D
multiwavelength photoacoustic images of vascular tissue by exploiting the unique spectral features of lipids.
Recognising regions of high lipid concentration would be useful to identify plaques which are likely to rupture
(vulnerable plaques). To investigate this, samples of post mortem human aortas were imaged at a range of
near-infrared (NIR) wavelengths and compared to histology. Photoacoustic images were also obtained when
illuminating the sample through blood. This study demonstrated that lipid rich atheromatous plaques can
clearly be identified using multiwavelength photoacoustic imaging.
Photoacoustic spectroscopy has been shown to be able to discriminate between normal and atheromatous areas
of arterial tissue in the visible range (410nm-680nm). However, at these wavelengths haemoglobin absorption is
also very high. This makes it challenging to apply photoacoustic techniques using an intravascular probe, as a
significant amount of the excitation light will be absorbed by the blood present in the artery. In this study we
investigate the use of a wider range of excitation wavelengths (740-1800nm) for discriminating between normal
arterial tissue and lipid rich plaques and minimise the effect of blood absorption. Special attention will be given
to the near infra-red (NIR) wavelength range (900-1300nm) as in this region blood absorption is relatively weak
and there are expected to be significant differences in the absorption spectrum of each tissue type. To investigate
this, tissue samples were obtained and imaged at a range of wavelengths, the samples were illuminated first
through water, then blood. This study demonstrated that the photoacoustic technique can discriminate between
normal arterial tissue and lipid rich plaques, even when blood is present.
KEYWORDS: Semiconductor lasers, Photoacoustic spectroscopy, Signal detection, Blood, Tissues, Signal generators, Laser systems engineering, Photoacoustic imaging, Imaging systems, Signal to noise ratio
Photoacoustic methods can be used to make spatially resolved spectroscopic measurements of blood oxygenation when using a multiwavelength excitation source, such as an OPO system. Since these excitation sources are usually expensive and bulky, an alternative is to use laser diodes. A fibre coupled laser diode excitation system has been developed, providing two wavelengths, 850 and 905nm, each composed of 6 high peak power pulsed laser diodes. The system provided variable pulse durations (65-500ns) and repetition rates of up to 5KHz. The pulse energies delivered by the excitation system at 905nm and 850nm were measured to be 120&mgr;J and 80&mgr;J respectively for a 200ns pulse duration. To demonstrate the utility of the system, the excitation source was combined with an ultrasound detector to form a probe for in vivo single point measurements of superficial blood vessels. Changes in blood oxygenation and volume in the finger tip were monitored while making venous and arterial occlusions. To demonstrate the imaging capability of the excitation system, 2D photoacoustic images of a physiologically realistic phantom were obtained for a range of pulse durations using a cylindrical scanning system. The phantom was composed of cylindrical absorbing elements (&mgr;a=1mm^{-1}) of 2.7mm diameter, immersed in a 1% intralipid solution (&mgr;s=1mm^{-1}). This study demonstrated the potential use of laser diodes as an excitation source for photoacoustic imaging of superficial vascular structures.
KEYWORDS: Semiconductor lasers, Photoacoustic spectroscopy, Signal to noise ratio, Signal detection, Signal generators, Signal attenuation, Acoustics, Biomedical optics, Capillaries, Laser systems engineering
An important capability of photoacoustic methods is the ability to make spatially resolved spectroscopic measurements
of blood oxygenation by imaging at multiple NIR excitation wavelengths, usually sourced from Q-switched
Nd:YAG pumped OPO based systems. These excitation sources are usually bulky, expensive and with limited
scope for varying the pulse repetition rate and pulse width. An alternative would be to use pulsed laser diodes
as excitation sources. To evaluate the possibility of developing a multiwavelength excitation system composed
of three wavelengths 810, 850 and 905nm, a single wavelength (905nm) system was built. To achieve a sufficient
SNR, four high peak power pulsed laser diodes were combined. The design of the laser drivers provided variable
pulse duration (65-500ns) and repetition rates of up to 2.5KHz. This allowed the pulse duration to be optimised
in order to (a) maximise the energy in the generated photoacoustic signal and (b) reduce the effects of frequency
dependent acoustic attenuation of tissue on the propagating acoustic wave by avoiding the generation of excessively
high frequency components. It also enabled the high repetition rate of laser diodes to be exploited in order
to average a large number of acquisitions over a short time period to increase SNR. Preliminary measurements
of SNR were made in phantoms using the single wavelength excitation system, to demonstrate the practical
biomedical utility of the system. A tissue phantom consisting of two capillaries (null set 460μm)filled with an absorbing
dye of similar optical properties to blood (μa ≈ 1mm-1), immersed at different depths in a 1% solution of
intralipid (μs ≈ 1mm-1) was used. To further demonstrate the capability of the system it was combined with a
cylindrical scanning system to image a strongly absorbing cylinder immersed to a depth of 1cm in 1% solution
of intralipid (μs ≈ 1mm-1). This study demonstrated the potential for using laser diodes as excitation sources
for pulsed photoacoustic spectroscopic biomedical applications.
KEYWORDS: Semiconductor lasers, Photoacoustic spectroscopy, Signal attenuation, Acoustics, Signal to noise ratio, Signal generators, Signal detection, Q switched lasers, Pulsed laser operation, Absorption
Photoacoustic signals are usually generated using bulky and expensive Q-switched Nd:YAG lasers, with limited scope for varying the pulse repetition frequency, wavelength and pulse width. An alternative would be to use laser diodes as excitation sources; these devices are compact, relatively inexpensive, and available in a wide variety of NIR wavelengths. Their pulse duration and repetition rates can also be varied arbitrarily enabling a wide range of time and frequency
domain excitation methods to be employed. The main difficulty to overcome when using laser diodes for pulsed photoacoustic excitation is their low peak power compared to Q-switched lasers. However, the much higher repetition rate of laser diodes (~ kHz) compared to many Q-switched laser systems (~ tens of Hz) enables a correspondingly greater number of events to be acquired and signal averaged over a fixed time period. This offers the prospect of significantly increasing the signal-to-noise ratio (SNR) of the detected photoacoustic signal. Choosing the wavelength of the laser diode to be lower than that of the water absorption peak at 940nm, may also provide a significant advantage over a system lasing at 1064nm for measurements in tissue. If the output of a number of laser
diodes is combined it then becomes possible, in principle, to obtain a SNR approaching that achievable with a Q-switched laser. It is also suggested that optimising the pulse duration of the laser diode may reduce the effects of frequency-dependent acoustic attenuation in tissue on the photoacoustic signal. To investigate this, a numerical model based on the Poisson solution to the wave equation was developed. To validate the model, a high peak power pulsed laser diode system was built. It was composed of a 905nm stacked array laser diode coupled to an optical fibre and driven by a high current laser diode driver. Measurements of the SNR of photoacoustic signals generated in a purely absorbing medium (ink) were made as a function of pulse duration. This preliminary study shows the potential for using laser diodes as excitation sources for photoacoustic applications in the biomedical field.
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