共查询到20条相似文献,搜索用时 31 毫秒
1.
Wang Y Maslov K Zhang Y Hu S Yang L Xia Y Liu J Wang LV 《Journal of biomedical optics》2011,16(1):011014
For broad applications in biomedical research involving functional dynamics and clinical studies, a photoacoustic microscopy system should be compact, stable, and fast. In this work, we use a fiber laser as the photoacoustic irradiation source to meet these goals. The laser system measures 45×56×13 cm3. The stability of the laser is attributed to the intrinsic optical fiber-based light amplification and output coupling. Its 50-kHz pulse repetition rate enables fast scanning or extensive signal averaging. At the laser wavelength of 1064 nm, the photoacoustic microscope still has enough sensitivity to image small blood vessels while providing high optical absorption contrast between melanin and hemoglobin. Label-free melanoma cells in flowing bovine blood are imaged in vitro, yielding measurements of both cell size and flow speed. 相似文献
2.
In order to image noninvasively cell nuclei in vivo without staining, we have developed ultraviolet photoacoustic microscopy (UV-PAM), in which ultraviolet light excites nucleic acids in cell nuclei to produce photoacoustic waves. Equipped with a tunable laser system, the UV-PAM was applied to in vivo imaging of cell nuclei in small animals. We found that 250 nm was the optimal wavelength for in vivo photoacoustic imaging of cell nuclei. The optimal wavelength enables UV-PAM to image cell nuclei using as little as 2 nJ laser pulse energy. Besides the optimal wavelength, application of a wavelength between 245 and 275 nm can produce in vivo images of cell nuclei with specific, positive, and high optical contrast. 相似文献
3.
We developed optical coherence photoacoustic microscopy (OC-PAM) to demonstrate that the functions of optical coherence tomography (OCT) and photoacoustic microscopy (PAM) can be achieved simultaneously by using a single illuminating light source. We used a pulsed broadband laser centered at 580 nm and detected the absorbed photons through photoacoustic detection and the back-scattered photons with an interferometer. In OC-PAM, each laser pulse generates both one OCT A-line and one PAM A-line simultaneously; as a result, the two imaging modalities are intrinsically co-registered in the lateral directions. In vivo images of the mouse ear were acquired to demonstrate the capabilities of OC-PAM. 相似文献
4.
Zhang C Maslov K Hu S Chen R Zhou Q Shung KK Wang LV 《Journal of biomedical optics》2012,17(2):020501
Submicron-resolution photoacoustic microscopy (PAM) currently exists only in transmission mode, due to the technical difficulties of combining high numerical-aperture (NA) optical illumination with high NA acoustic detection. The lateral resolution of reflection-mode PAM has not reached <2 μm in the visible light range. Here we develop the first reflection-mode submicron-resolution PAM system with a new compact design. By using a parabolic mirror to focus and reflect the photoacoustic waves, sufficient signals were collected for good sensitivity without distorting the optical focusing. By imaging nanospheres and a resolution test chart, the lateral resolution was measured to be ≈ 0.5 μm with an optical wavelength of 532 nm, an optical NA of 0.63. The axial resolution was measured at 15 μm. Here the axial resolution was measured by a different experiment with the lateral resolution measurement. But we didn't describe the details of axial resolution measurement due to space limit. The maximum penetration was measured at ≈ 0.42 mm in optical-scattering soft tissue. As a comparison, both the submicron-resolution PAM and a 2.4 μm-resolution PAM were used to image a mouse ear in vivo with the same optical wavelength and similar pulse energy. Capillaries were resolved better by the submicron-resolution PAM. Therefore, the submicron-resolution PAM is suitable for in vivo high-resolution imaging, or even subcellular imaging, of optical absorption. 相似文献
5.
Both photoacoustic imaging and power Doppler ultrasound are capable of producing images of the vasculature of living subjects, however, the contrast mechanisms of the two modalities are very different. We present a quantitative and objective comparison of the two methods using phantom data, highlighting relative merits and shortcomings. An imaging system for combined photoacoustic and high-frequency power Doppler ultrasound microscopy is presented. This system uses a swept-scan 25-MHz ultrasound transducer with confocal dark-field laser illumination optics. A pulse-sequencer enables ultrasonic and laser pulses to be interlaced so that photoacoustic and power Doppler ultrasound images can be coregistered. Experiments are performed on flow phantoms with various combinations of vessel size, flow velocity, and optical wavelength. For the task of blood volume detection, power Doppler is seen to be advantageous for large vessels and high flow speeds. For small vessels with low flow speeds, photoacoustic imaging is seen to be more effective than power Doppler at the detection of blood as quantified by receiver operating characteristic analysis. A combination of the two modes could provide improved estimates of fractional blood volume in comparison with either mode used alone. 相似文献
6.
Shi W Kerr S Utkin I Ranasinghesagara J Pan L Godwal Y Zemp RJ Fedosejevs R 《Journal of biomedical optics》2010,15(5):056017
Optical-resolution photoacoustic microscopy (OR-PAM) is a novel imaging technology for visualizing optically absorbing superficial structures in vivo with lateral spatial resolution determined by optical focusing rather than acoustic detection. Since scanning of the illumination spot is required, OR-PAM imaging speed is limited by both scanning speed and laser pulse repetition rate. Unfortunately, lasers with high repetition rates and suitable pulse durations and energies are not widely available and can be cost-prohibitive and bulky. We are developing compact, passively Q-switched fiber and microchip laser sources for this application. The properties of these lasers are discussed, and pulse repetition rates up to 100 kHz are demonstrated. OR-PAM imaging was conducted using a previously developed photoacoustic probe, which enabled flexible scanning of the focused output of the lasers. Phantom studies demonstrate the ability to image with lateral spatial resolution of 7±2 μm with the microchip laser system and 15±5 μm with the fiber laser system. We believe that the high pulse repetition rates and the potentially compact and fiber-coupled nature of these lasers will prove important for clinical imaging applications where real-time imaging performance is essential. 相似文献
7.
We describe the phenomenon of a sudden decrease in the amplitude of photoacoustic signals arising from nanosecond laser pulse irradiation of biological samples, measured in vitro. Several dental enamel and chicken/turkey breast samples are examined. Moderate optical energy densities (i.e., about 300 mJ/cm2) are used, typical of those exploited in photoacoustic investigations. Measurements show a rapid decay of photoacoustic signals within the first few laser pulses absorbed by the sample. This phenomenon indicates that laser irradiation interacts with biological samples, causing long-term physical changes that can be attributed to a reduction of optical absorption within the samples. 相似文献
8.
Pulsed photoacoustic spectroscopy was used to measure blood oxygen saturation in vitro. An optical parametric oscillator laser system provided nanosecond excitation pulses over the wavelength range 740-1040 nm which were used to generate photoacoustic signals in a cuvette through which a saline suspension of red blood cells was circulated. The signal amplitude and the effective attenuation coefficient were extracted from the photoacoustic signals as a function of wavelength to provide photoacoustic spectra of the blood. From these, the relative concentrations of oxy- and deoxyhaemoglobin, and therefore blood oxygen saturation (SO2), were determined using forward models of the absorbed energy distribution based on diffusion theory. A standard linear model of the dependence of absorbance on the concentration of chromophores was also used to calculate the blood oxygen saturation from the signal amplitude spectra. The diffusion approximation model was shown to produce the highest accuracy in blood SO2. The photoacoustically determined oxygen saturation was found to have an accuracy of +/-4% SO2 for signal amplitude data and +/-2.5% SO2 for effective attenuation spectra. The smallest change in oxygen saturation that can be measured using this technique was +/-1% SO2. 相似文献
9.
Sivaramakrishnan M Maslov K Zhang HF Stoica G Wang LV 《Physics in medicine and biology》2007,52(5):1349-1361
We investigate the feasibility of obtaining accurate quantitative information, such as local blood oxygenation level (sO2), with a spatial resolution of about 50 microm from spectral photoacoustic (PA) measurements. The optical wavelength dependence of the peak values of the PA signals is utilized to obtain the local blood oxygenation level. In our in vitro experimental models, the PA signal amplitude is found to be linearly proportional to the blood optical absorption coefficient when using ultrasonic transducers with central frequencies high enough such that the ultrasonic wavelengths are shorter than the light penetration depth into the blood vessels. For an optical wavelength in the 578-596 nm region, with a transducer central frequency that is above 25 MHz, the sensitivity and accuracy of sO2 inversion is shown to be better than 4%. The effect of the transducer focal position on the accuracy of quantifying blood oxygenation is found to be negligible. In vivo oxygenation measurements of rat skin microvasculature yield results consistent with those from in vitro studies, although factors specific to in vivo measurements, such as the spectral dependence of tissue optical attenuation, dramatically affect the accuracy of sO2 quantification in vivo. 相似文献
10.
The inverse relationship between two-photon excited fluorescence (TPEF) and laser pulse duration suggests that two-photon microscopy (TPM) performance may be improved by decreasing pulse duration. However, for ultrashort pulses of sub-10 femtosecond (fs) in duration, its spectrum contains the effective gain bandwidth of Ti:Sapphire and its central wavelength is no longer tunable. An experimental study was performed to explore this apparent tradeoff between untuned sub-10 fs transform-limited pulse (TLP) and tunable 140 fs pulse for TPEF. Enhancement factors of 1.6, 6.7, and 5.2 are measured for Indo-1, FITC, and TRITC excited by sub-10 fs TLP compared with 140 fs pulse tuned to the two-photon excitation (TPE) maxima at 730 nm, 800 nm, and 840 nm, respectively. Both degenerate (v(1) = v(2)) and nondegenerate (v(1) ≠ v(2)) mixing of sub-10 fs TLP spectral components result in its broad second-harmonic (SH) power spectrum and high spectral density, which can effectively compensate for the lack of central wavelength tuning and lead to large overlap with dye TPE spectra for TPEF enhancements. These pulse properties were also exploited for demonstrating its potential applications in multicolor imaging with TPM. 相似文献
11.
A method is proposed that utilizes the advantages of optical ultrasound detection in two-dimensional photoacoustic section imaging, combining an optical interferometer with an acoustic mirror. The concave mirror has the shape of an elliptical cylinder and concentrates the acoustic wave generated around one focal line in the other one, where an optical beam probes the temporal evolution of acoustic pressure. This yields line projections of the acoustic sources at distances corresponding to the time of flight, which, after rotating the sample about an axis perpendicular to the optical detector, allows reconstruction of a section using the inverse Radon transform. A resolution of 120 [micro sign]m within and 1.5 mm between the sections can be obtained with the setup. Compared to a bare optical probe beam, the signal-to-noise ratio (SNR) is seven times higher with the mirror. Furthermore, the imaging system is tested on a biological sample. 相似文献
12.
A fiber-based multimodal imaging system that combines multiphoton microscopy (MPM) with Fourier domain optical coherence microscopy (OCM) is reported. The system uses a fiber-based femtosecond laser, a fiber coupler, and a double-clad fiber (DCF) device. The fiber laser has a central wavelength of 1.04 μm and bandwidth of 29 nm. Longer excitation wavelength is used to increase penetration depth and increase the excitation efficiency for dyes, such as red fluorescent dyes. A single mode fiber coupler is used to replace the free-space beam splitter and one arm of the coupler is fused with a double-clad fiber device. The MPM and OCM share the same excitation light path in the core of a double-clad fiber, while the OCM and MPM signals were collected by the core and clad of the double-clad fiber, respectively. The performance of the introduced double-clad device is analyzed. The device can confine all the excitation light in the core and has a collection efficiency of 20% for the MPM signal. The efficiency can be further increased by fusing more multimode fibers with the DCF. Simultaneous optical coherence microscopic imaging, second harmonic generation imaging, and two-photon excitation fluorescence imaging are demonstrated in biological samples. 相似文献
13.
We demonstrate the use of tone-burst excitation and time-gated spectral analysis for photoacoustic Doppler mapping of flow in an unperturbed vessel phantom and in a vessel with a spatially varying lumen. The method, which mimics pulsed Doppler ultrasound, enables simultaneous measurement of axial position and flow as well as complete characterization of the Doppler spectrum over a wide range of mean velocities (3.5 to 200 mm∕s). To generate the required optical excitation, a continuous cw laser source followed by an external electro-optic modulator is used. Stenoses at various levels are emulated in a C-flex tube with a flowing suspension of micrometer-scale carbon particles. Two-dimensional maps of spectral content versus axial position at different points along the vessel and for various levels of perturbations demonstrate the potential use of the method for characterization of flow irregularities. 相似文献
14.
Laser photoacoustic determination of physiological glucose concentrations in human whole blood 总被引:3,自引:0,他引:3
A glucose concentration analysis of human whole blood samples has been accomplished using pulsed laser photoacoustic spectroscopy
(LPAS). Using a CO2 laser operating with μJ pulse energies, the technique has shown the required discrimination and sensitivity to determine
glucose concentrations within the physiological range (18–450 mg dl−1) in whole blood samples. The sensitivity achieved with this system is comparable to that of the existing commerical enzyme-based
diagnostic systems presently used in hospital clinical chemistry environments. The technique is compared with other optical
methods that have recently been used for glucose determination, and its applicability for use in the development of an in
vivo monitor is discussed. 相似文献
15.
A quantitative flow measurement method that utilizes a sequence of photoacoustic images is described. The method is based on the use of gold nanorods as a contrast agent for photoacoustic imaging. The peak optical absorption wavelength of a gold nanorod depends on its aspect ratio, which can be altered by laser irradiation (we establish a wash-in flow estimation method of this process). The concentration of nanorods with a particular aspect ratio inside a region of interest is affected by both laser-induced shape changes and replenishment of nanorods at a rate determined by the flow velocity. In this study, the concentration is monitored using a custom-designed, high-frame-rate photoacoustic imaging system. This imaging system consists of fiber bundles for wide area laser irradiation, a laser ultrasonic transducer array, and an ultrasound front-end subsystem that allows acoustic data to be acquired simultaneously from 64 transducer elements. Currently, the frame rate of this system is limited by the pulse-repetition frequency of the laser (i.e., 15 Hz). With this system, experimental results from a chicken breast tissue show that flow velocities from 0.125 to 2 mms can be measured with an average error of 31.3%. 相似文献
16.
We demonstrate a compact and self-starting fiber-delivered femtosecond Cr:forsterite laser for nonlinear light microscopy. A semiconductor saturable absorber mirror provides the self-starting mechanism and maintains long-term stability in the laser cavity. Four double-chirped mirrors are employed to reduce the size of the cavity and to compensate for group velocity dispersion. Delivered by a large-mode-area photonic crystal fiber, the generated laser pulses can be compressed down to be with a nearly transform-limited pulse width with 2.2-nJ fiber-output pulse energy. Based on this fiber-delivered Cr:forsterite laser source, a compact and reliable two-photon fluorescence microscopy system can thus be realized. 相似文献
17.
High-energy and short-duration laser pulses are desirable to improve the photoacoustic image quality when imaging deeply seated lesions. In many clinical applications, the high-energy pulses are coupled to tissue using optical fibers. These pulses can damage fibers if the damage threshold is exceeded. While keeping the total energy under the Food and Drug Administration limit for avoiding tissue damage, it is necessary to reduce the peak intensity and increase the pulse duration for minimizing fiber damage and delivering sufficient light for imaging. We use laser-pulse-stretching to address this problem. An initial 17-ns pulse was stretched to 27 and 37 ns by a ring-cavity laser-pulse-stretching system. The peak power of the 37-ns stretched pulse reduced to 42% of the original, while the fiber damage threshold was increased by 1.5-fold. Three ultrasound transducers centered at 1.3-, 3.5-, and 6-MHz frequencies were simulated, and the results showed that the photoacoustic signal of a 0.5-mm-diameter target obtained with 37-ns pulse was about 98, 91, and 80%, respectively, using the same energy as the 17-ns pulse. Simulations were validated using a broadband hydrophone. Quantitative comparisons of photoacoustic images obtained with three corresponding transducers showed that the image quality was not affected by stretching the pulse. 相似文献
18.
McConnell G 《Physics in medicine and biology》2007,52(3):717-724
A singly resonant femtosecond-pulsed synchronously pumped optical parametric oscillator (SPOPO) is reported that provides a wavelength-tunable alternative to the Cr:forsterite source for nonlinear microscopy applications. Periodically poled lithium niobate within an external resonator was synchronously pumped by a commercial fs-pulsed Nd:YLF laser source to provide fs-pulsed continuously tunable radiation from 1.45 to 1.63 microm. Third harmonic generation (THG) microscopy and three-photon laser scanning fluorescence microscopy (3PLSM) of a range of samples were subsequently performed using this long-wavelength SPOPO source. The ability to tune the excitation wavelength delivered to the sample significantly improved the efficiency of both the 3PLSM and THG imaging experiments. 相似文献
19.
Our aim is the development of a simple optical method for pulse wave profile, pulse wave delay time, and blood flow measurement. The method is based on recording the Doppler frequency shift related to a moving target--blood vessel walls or small particles. The Doppler signal is detected using the self-mixing that occurs in the diode laser cavity when radiation scatters back from the moving target into the laser and interferes with the field inside. Two different ways can be simultaneously used for the self-mixing signal extraction: a photodiode accommodated in the rear facet of the diode laser package and a resistor from the laser pump current. An experimental device with a pigtail laser diode is developed that is able to detect the pulsation of major arteries with potentially useful information, including the pulse wave profile and the pulse wave delay time. The pulse wave delay time in different regions of the human body is measured relative to the electrocardiogram (ECG) signal. Also the flow velocity of a liquid suspension containing particles the size of erythrocytes (equivalent to blood flow) is measured. Registered signals are stored after digitalization and preprocessed using LabView for a Windows environment. The described device has the application of the self-mixing method and highlights significant advantages of simplicity, compactness, and robustness as well as the self-aligning and self-detecting abilities of such method, compared with the use of conventional interferometric method. 相似文献
20.
Fluorescence tomography (FT) reconstructs the three-dimensional (3D) fluorescent reporter probe distribution inside biological tissue. These probes target molecules of biological function, e.g. cell surface receptors or enzymes, and emit fluorescence light upon illumination with an external light source. The fluorescence light is detected on the tissue surface and a source reconstruction algorithm based on the simplified spherical harmonics (SP(N)) equations calculates the unknown 3D probe distribution inside tissue. While current FT approaches require multiple external sources at a defined wavelength range, the proposed FT method uses only a white light source with tunable wavelength selection for fluorescence stimulation and further exploits the spectral dependence of tissue absorption for the purpose of 3D tomographic reconstruction. We will show the feasibility of the proposed hyperspectral excitation-resolved fluorescence tomography method with experimental data. In addition, we will demonstrate the performance and limitations of such a method under ideal and controlled conditions by means of a digital mouse model and synthetic measurement data. Moreover, we will address issues regarding the required amount of wavelength intervals for fluorescent source reconstruction. We will explore the impact of assumed spatially uniform and nonuniform optical parameter maps on the accuracy of the fluorescence source reconstruction. Last, we propose a spectral re-scaling method for overcoming the observed limitations in reconstructing accurate source distributions in optically non-uniform tissue when assuming only uniform optical property maps for the source reconstruction process. 相似文献