Optical-resolution photoacoustic microscopy (OR-PAM) has demonstrated the capability of mapping microvasculature and is promising for biomedical applications. However, the popularity of the PAM system is limited due to the use of expansive and bulky laser sources. In this paper, a compact and cost-efficient OR-PAM system with a laser diode excitation has been developed. The black threads, phantom made of polyethylene tubes filled with rat blood, and a mouse ear were imaged to evaluate the PAM and demonstrate its capability of imaging biological tissue.
We have estimated the micro-mechanical properties of ovarian tissue using phase-sensitive swept source optical coherence tomography. Ovary samples were mechanically excited by periodical vibration of an ultrasound transducer. The displacement and strain of the tissues were calculated during loading. Significant difference in strain was observed between the normal and malignant ovary groups, which indicates much softer and heterogeneous tissue structure for malignant ovaries. The initial results show that the phase sensitive swept source optical coherence elastography (OCE) can be an effective tool for characterization of stiffness and other micro-mechanical properties of normal and malignant ovarian tissue.
In this paper, human ovarian tissues with malignant and benign features were imaged ex vivo by using an opticalresolution photoacoustic microscopy (OR-PAM) system. Several features were quantitatively extracted from PAM images to describe photoacoustic signal distributions and fluctuations. 106 PAM images from 18 human ovaries were classified by applying those extracted features to a logistic prediction model. 57 images from 9 ovaries were used as a training set to train the logistic model, and 49 images from another 9 ovaries were used to test our prediction model. We assumed that if one image from one malignant ovary was classified as malignant, it is sufficient to classify this ovary as malignant. For the training set, we achieved 100% sensitivity and 83.3% specificity; for testing set, we achieved 100% sensitivity and 66.7% specificity. These preliminary results demonstrate that PAM could be extremely valuable in assisting and guiding surgeons for in vivo evaluation of ovarian tissue.
A high-throughput ultrasound/photoacoustic probe for delivering high contrast and signal-to-noise ratio images was designed, constructed, and tested. The probe consists of a transvaginal ultrasound array integrated with four 1mm-core optical fibers and a sheath. The sheath encases transducer and is lined with highly reflecting aluminum for high intensity light output and uniformity while at the same time remaining below the maximum permissible exposure (MPE) recommended by the American National Standards Institute (ANSI). The probe design was optimized by simulating the light fluence distribution in Zemax. The performance of the probe was evaluated by experimental measurements of the fluence and real-time imaging of polyethylene-tubing filled with blood. These results suggest that our probe has great potential for in vivo imaging and characterization of ovarian cancer.
Tumor hypoxia is a major indicator of treatment resistance to chemotherapeutic drugs, and fluorescence optical tomography has tremendous potential to provide clinically useful, functional information by identifying tumor hypoxia. The synthesis of a 2-nitroimidazole-indocyanine green conjugate using a piperazine linker (piperazine-2-nitroimidazole-ICG) capable of robust fluorescent imaging of tumor hypoxia is described. In vivo mouse tumor imaging studies were completed and demonstrate an improved imaging capability of the new dye relative to an earlier version of the dye that was synthesized with an ethanolamine linker (ethanolamine-2-nitroimidazole-ICG). Mouse tumors located at imaging depths of 1.5 and 2.0 cm in a turbid medium were imaged at various time points after intravenous injection of the dyes. On average, the reconstructed maximum fluorescence concentration of the tumors injected with piperazine-2-nitroimidazole-ICG was twofold higher than that injected with ethanolamine-2-nitroimidazole-ICG within 3 h postinjection period and 1.6 to 1.7 times higher beyond 3 h postinjection. The untargeted bis-carboxylic acid ICG completely washed out after 3 h postinjection. Thus, the optimal window to assess tumor hypoxia is beyond 3 h postinjection. These findings were supported with fluorescence images of histological sections of tumor samples and an immunohistochemistry technique for identifying tumor hypoxia.
In this paper, we have synthesized a second generation tumor hypoxia targeted 2-nitroimidazole-ICG conjugate using piperazine linker (2-nitro-ICG-p) and validated its performance in in vivo tumor targeting. The results have shown that tumor hypoxia can be targeted with twice higher signal strength beyond three hours post-injection while the un-targeted ICG has completely washed out. The improvement of the second generation 2-nitro-ICG-p dyes is 1.2-1.3 times over the first generation 2-nitro-ICG dyes using ethanol linker beyond 3 hours post-injection which is the optimal time-window for evaluating tumor hypoxia.
We characterized ovarian tissue by using polarization-sensitive optical coherence tomography (PS-OCT). Phase retardation changing rate along with optical scattering coefficient and phase retardation were extracted from 33 ex vivo human ovaries from 18 patients. By using phase retardation changing rate as a classifier, we could achieve 71% sensitivity and 100% specificity. Combining those three parameters together, we could achieve 100% sensitivity and 100% specificity. These initial results showed that the quantitative analysis of PS-OCT could be a useful tool to characterize normal and malignant ovarian tissue.
Angle-resolved optical scattering properties of ovarian tissue on different optical coherence tomography (OCT) imaging planes were quantitatively measured by fitting the compounded OCT A-lines into a single scattering model. Higher cross correlation value of angle-resolved scattering coefficients between different OCT imaging planes was found in normal ovaries than was present in malignant ovaries. The mean cross correlation coefficient (MCC) was introduced in this pilot study to characterize and differentiate normal and malignant ovaries. A specificity of 100% and a sensitivity of 100% were achieved by setting MCC threshold at 0.6 in the limited sample population. The collagen properties such as content, structure and directivity were found to be different within OCT imaging penetration depth between normal and malignant ovarian tissue. The homogeneous three-dimensional collagen fiber network observed in the normal ovary effectively explains the stronger cross correlation of angle-resolved scattering properties on different imaging planes while the heterogeneity observed in the malignant ovary suggests a weaker correlation.
In this paper, we present the construction of an optical-resolution photoacoustic microscopy (OR-PAM) system and
studies done on the characterization of human ovarian tissue with malignant and benign features ex vivo. PAM images of
the ovaries showed more detailed blood vessel distributions with much higher resolution compared with conventional
photoacoustic images obtained with array transducers. In all, 29 PAM images (20 from normal ovaries and 9 from
malignant ovaries) were studied. Eight different features were extracted quantitatively from the PAM images, and a
generalized linear model (GLM) was used to classify the ovaries as normal or malignant. By using the GLM, a
specificity of 100% and a sensitivity of 100% were obtained for the training set. These preliminary results demonstrate
the feasibility of our PAM system in mapping microvasculature networks, as well as characterizing the ovarian tissue,
and could be extremely valuable in assisting surgeons for in vivo evaluation of ovarian tissue.
Angle-resolved optical scattering properties of ovarian tissue, on different optical coherence tomography (OCT) imaging planes, were quantitatively measured by fitting the compounded OCT A-lines into a single scattering model. Higher cross correlation value of angle-resolved scattering coefficients between different OCT imaging planes was found in normal ovaries than was present in malignant ovaries. The mean cross correlation coefficient (MCC) was introduced in this pilot study to characterize and differentiate normal, n=6 , and malignant, n=4 , ovaries. A specificity of 100 percent and a sensitivity of 100 percent were achieved by setting MCC threshold at 0.6. Collagen properties, within the OCT imaging penetration depth, were also qualitatively studied in terms of their content, structure and directivity. The homogeneous three-dimensional collagen fiber network, observed in the normal ovary, effectively explains the stronger cross correlation of angle-resolved scattering properties on different imaging planes while the heterogeneity, observed in the malignant ovary, suggests a weaker correlation.
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.
High-energy and short-duration outputs from lasers are desirable to improve the photoacoustic image quality when
imaging deeply-seated lesions. In many clinical applications, optical fibers are used to couple the high-energy laser pulse
to tissue. These high peak intensity pulses can damage an optical fiber input face if the damage threshold is exceeded. It
is necessary to reduce the peak intensity to minimize the fiber damage and to delivery sufficient light for imaging. In this
paper, a laser-pulse-stretching technique is introduced to reduce the peak intensity of laser pulses. To demonstrate the
technique, an initial 17ns pulse was stretched to 37ns by a ring-cavity laser-pulse-stretching system, and the laser peak
power reduced to 42%. The stretched pulse increased the fiber damage threshold by 1.5-fold. Three ultrasound
transducers centered at 1.3MHz, 3.5MHz, 6MHz frequencies were simulated and the results showed that the
photoacoustic signal of 0.5mm-diameter target obtained with 37ns pulse was about 98%, 91% and 80% respectively
using the same energy as with the 17ns pulse. Simulations were validated using a broadband hydrophone. Quantitative
comparisons of photoacoustic images obtained with three corresponding ultrasound transducers showed that the image
quality was not affected by stretching the pulse.
In this paper, we report an intraoperative approach by combining optical coherence tomography (OCT) and position
detection to detect and characterize ovarian cancers. A total of 18 ovaries were studied ex vivo. Based on histopathology
result, they were classified into normal and malignant groups, respectively. On average positron count rate of 8.0-fold
higher was found between malignant and normal ovaries. OCT imaging of ovaries revealed many detailed morphologic
features that could be potentially valuable for detecting early malignant changes in ovarian tissue. Optical scattering
coefficients of these ovaries were estimated from OCT A-lines. Normal ovarian tissue showed higher scattering
coefficient than that of malignant ovarian tissue. Using a threshold of 2.00 mm-1 for all ovaries, a sensitivity of 100% and a
specificity of 100% were achieved. This initial data shows our intraoperative probe based on OCT and positron detection
has a great potential for ovarian cancer detection and characterization.
Optical scattering coefficient from ex-vivo unfixed normal and malignant ovarian tissue was quantitatively extracted by
fitting optical coherence tomography (OCT) A-line signals to a single scattering model. 1097 average A-line
measurements at a wavelength of 1310nm were performed at 108 sites obtained from 18 ovaries. The average scattering
coefficient obtained from normal group consisted of 833 measurements from 88 sites was 2.41 mm-1 (±0.59), while the
average coefficient obtained from malignant group consisted of 264 measurements from 20 sites was 1.55 mm-1 (±0.46).
Using a threshold of 2 mm-1 for each ovary, a sensitivity of 100% and a specificity of 100% were achieved. The amount of
collagen within OCT imaging depth was analyzed from the tissue histological section stained with Sirius Red. The average
collagen area fraction (CAF) obtained from normal group was 48.4% (±12.3%), while the average CAF obtained from
malignant group was 11.4% (±4.7%). Statistical significance of the collagen content was found between the two groups
(p < 0.001). The preliminary data demonstrated that quantitative extraction of optical scattering coefficient from OCT
images could be a potential powerful method for ovarian cancer detection and diagnosis.
Optical scattering coefficient from ex vivo unfixed normal and malignant ovarian tissue was quantitatively extracted by fitting optical coherence tomography (OCT) A-line signals to a single scattering model. 1097 average A-line measurements at a wavelength of 1310 nm were performed at 108 sites obtained from 18 ovaries. The average scattering coefficient obtained from the normal tissue group consisted of 833 measurements from 88 sites was 2.41 mm−1 (±0.59), while the average coefficient obtained from the malignant tissue group consisted of 264 measurements from 20 sites was 1.55 mm−1 (±0.46). The malignant ovarian tissue showed significant lower scattering than the normal group (p < 0.001). The amount of collagen within OCT imaging depth was analyzed from the tissue histological section stained with Sirius Red. The average collagen area fraction (CAF) obtained from the normal tissue group was 48.4% (±12.3%), while the average CAF obtained from the malignant tissue group was 11.4% (±4.7%). A statistical significance of the collagen content was found between the two groups (p < 0.001). These results demonstrated that quantitative measurements of optical scattering coefficient from OCT images could be a potential powerful method for ovarian cancer detection.
Photoacoustic imaging and optical coherence tomography are two emerging imaging modalities which provide
complementary optical absorption and scattering contrasts for biological tissues. While photoacoustic imaging provides
tissue vasculature information, optical coherence tomography offers micron-scale morphological imaging with
penetration depths of 1~3 mms. Pulse-echo ultrasound is readily available from photoacoustic system and it provides
tissue structure information at deeper depths with resolutions scalable with the transducer frequency. We present a
prototype endoscope that consists of a ball lensed optical coherence tomography probe, a right-angled multimode fiber
for delivering the laser beam for photoacoustic imaging, and a high frequency ultrasound transducer of 35 MHz center
frequency. The overall diameter is 5mms. Porcine ovaries were imaged ex vivo to demonstrate the capability of this new
combined endoscopy. The microvascular and high resolution structural images at subsurface and deeper tissue range
demonstrate the synergy of the combined endoscopy over each modality alone.
A fiber optical coherence tomography (OCT) probe is used for three dimensional dental imaging. The probe has a
lightweight miniaturized design with a size of a pen to facilitate clinic in vivo diagnostics. The probe is interfaced with a
swept-source / Fourier domain optical coherence tomography at 20K axial scanning rate. The tooth samples were
scanned from occlusal, buccal, lingual, mesial, and distal orientations. Three dimensional imaging covers tooth surface
area up to 10 mm x 10 mm with a depth about 5 mm, where a majority of caries affection occurs. OCT image provides
better resolution and contrast compared to gold standard dental radiography (X-ray). In particular, the technology is well
suited for occlusal caries detection. This is complementary to X-ray as occlusal caries affection is difficult to be detected
due to the X-ray projectile scan geometry. The 3D topology of occlusal surface as well as the dentin-enamel junction
(DEJ) surface inside the tooth can be visualized. The lesion area appears with much stronger back scattering signal
intensity.
Ovarian cancer has the lowest survival rate of the gynecologic cancers with a 5-year survival of about 50% in the United
States. With current screening and diagnostic abilities for ovarian cancers, most of the diagnosed patients are already with
advanced stages and the majority of them will die of this deadly disease. In this paper, we report a multimodal imaging
approach which combines optical coherence tomography (OCT) and positron detection for early ovarian cancer detection.
The dual modality system has the capability of providing both functional and morphological images simultaneously. While
the positron detection provides the metabolism activity of the ovary due to the uptake of radiotracer, the OCT provides the
high resolution (25μm X 25μm X 12μm - longitudinal X lateral X axial in air) structural imaging at 20k A-lines per second.
Total 18 ovaries obtained from 10 patients classified as normal, abnormal and malignant ovarian tissues were characterized
ex vivo. Positron counts of 1.2-fold higher was found between abnormal and normal ovaries and 3~30-fold higher was
found between malignant and normal ovaries. OCT imaging of malignant and abnormal ovaries revealed many detailed
morphologic features that could be potentially valuable for detecting early malignant changes in the ovary.
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