Non-telecentric photoacoustic microscopy for multi-scale imaging.

Optical-resolution photoacoustic microscopy (OR-PAM) excels in precisely imaging a biological tissue based on absorption contrast. However, existing OR-PAMs are confined by fixed compromises between spatial resolution and field of view (FOV), preventing the integration of large FOV and local high-resolution within one system. Here, we present a non-telecentric OR-PAM (nTC-PAM) that empowers efficient adaptation of FOV and spatial resolution to match the multi-scale requirement of diverse biological imaging. Our method allows for a large-scale transformation in FOV and even surpassing the nominal FOV of the objective with minimal marginal degradation of the lateral resolution. We demonstrate the advantage of nTC-PAM through multi-scale imaging of the leaf phantom, mouse ear, and cortex. The results reveal that nTC-PAM can switch the FOV and spatial resolution to meet the requirements of different biological tissues, such as large-scale imaging of the whole cerebral cortex and high-resolution imaging of microvascular structures in local brain regions.

Real-time mapping of photo-sono therapy induced cavitation using Doppler optical coherence tomography.

Photo-sono therapy (PST) is an innovative anti-vascular approach based on cavitation-induced spallation. Currently, passive cavitation detection (PCD) is the prevalent technique for cavitation monitoring during treatment. However, the limitations of PCD are the lack of spatial information of bubbles and the difficulty of integration with the PST system. To address this, we proposed a new, to the best of our knowledge, cavitation mapping method that integrates Doppler optical coherence tomography (OCT) with PST to visualize bubble dynamics in real time. The feasibility of the proposed system has been confirmed through experiments on vascular-mimicking phantoms and in vivo rabbit ear vessels, and the results are compared to high-speed camera observations and PCD data. The findings demonstrate that Doppler OCT effectively maps cavitation in real time and holds promise for guiding PST treatments and other cavitation-related clinical applications.

Miniaturized preamplifier integration in ultrasound transducer design for enhanced photoacoustic imaging.

Photoacoustic imaging (PAI) utilizes the photoacoustic effect to record both vascular and functional characteristics of a biological tissue. Photoacoustic signals have typically low amplitude that cannot be read efficiently by data acquisition systems. This necessitates the use of one or more amplifiers. These amplifiers are somewhat bulky (e.g., the ZFL-500LN+, Mini-Circuits, USA, or 351A-3-50-NI, Analog Modules Inc., USA). Here, we describe the fabrication and development process of a transducer with a built-in low-noise preamplifier that is encased within the transducer housing. This new, to the best of our knowledge, design could be advantageous for applications where a compact transducer + preamplifier is required. We demonstrate the performance of this compact detection unit in a laser scanning photoacoustic microscopy system by imaging a rat ear ex vivo and a rat brain vasculature in vivo.

Improvement of spatial resolution in photoacoustic microscopy using transmissive adaptive optics with a low-frequency ultrasound transducer.

Maintaining a high spatial resolution in photoacoustic microscopy (PAM) of deep tissues is difficult due to large aberration in an objective lens with high numerical aperture and photoacoustic wave attenuation. To address the issue, we integrate transmission-type adaptive optics (AO) in high-resolution PAM with a low-frequency ultrasound transducer (UT), which increases the photoacoustic wave detection efficiency. AO improves lateral resolution and depth discrimination in PAM, even for low-frequency ultrasound waves by focusing a beam spot in deep tissues. Using the proposed PAM, we increased the lateral resolution and depth discrimination for blood vessels in mouse ears.

3D printing of self-standing and vascular supportive multimaterial hydrogel structures for organ engineering.

Three dimensional printable formulation of self-standing and vascular-supportive structures using multi-materials suitable for organ engineering is of great importance and highly challengeable, but, it could advance the 3D printing scenario from printable shape to functional unit of human body. In this study, the authors report a 3D printable formulation of such self-standing and vascular-supportive structures using an in-house formulated multi-material combination of albumen/alginate/gelatin-based hydrogel. The rheological properties and relaxation behavior of hydrogels were analyzed before the printing process. The suitability of the hydrogel in 3D printing of various customizable and self-standing structures, including a human ear model, was examined by extrusion-based 3D printing. The structural, mechanical, and physicochemical properties of the printed scaffolds were studied systematically. Results supported the 3D printability of the formulated hydrogel with self-standing structures, which are customizable to a specific need. In vitro cell experiment showed that the formulated hydrogel has excellent biocompatibility and vascular supportive behavior with the extent of endothelial sprout formation when tested with human umbilical vein endothelial cells. In conclusion, the present study demonstrated the suitability of the extrusion-based 3D printing technique for manufacturing complex shapes and structures using multi-materials with high fidelity, which have great potential in organ engineering.

Photoacoustic-fluorescence microendoscopy in vivo.

A miniature endoscope capable of imaging multiple tissue contrasts in high resolution is highly attractive, because it can provide complementary and detailed tissue information of internal organs. Here we present a photoacoustic (PA)-fluorescence (FL) endoscope for optical-resolution PA microscopy (PAM) and FL microscopy (FLM). The endoscope with a diameter of 2.8 mm achieves high lateral resolutions of 5.5 and 6.3 µm for PAM and FLM modes, respectively. In vivo imaging of zebrafish larvae and a mouse ear is conducted, and high-quality images are obtained. Additionally, in vivo endoscopic imaging of a rat rectum is demonstrated, showing the endoscopic imaging capability of our endoscope. By providing dual contrasts with high resolution, the endoscope may open up new opportunities for clinical endoscopic imaging applications.

Cerebral angiography using transauricular access in a rabbit model: a new technique.

BACKGROUND: Cerebral angiography in a rabbit model is widely used in the field of interventional radiology. Conventionally, the femoral artery is used for cerebral angiography in radiology departments. However, angiographic studies require surgical cutdown of the femoral artery, which is technically difficult. PURPOSE: To evaluate a new cerebral angiography technique involving a transauricular approach in a rabbit model. MATERIAL AND METHODS: In each of 10 rabbits, central auricular arteries were punctured in the right or left ear with a 20-gauge i.v. catheter. A microcatheter (2.0 F) with a 0.016-inch guide wire was introduced through the i.v. catheter and advanced to the aortic arch. The microcatheter and guide wire were advanced selectively into cerebral arteries and angiography was performed. RESULTS: Central auricular arteries were successfully punctured with 20-gauge i.v. catheters. After approaching the aortic arch, microcatheter tips and guide wires were advanced manually to cerebral arteries on both sides. Difficulties in selecting the carotid arteries were resolved by using a looping technique within the cardiac chamber. Microcatheter loops within the cardiac chamber disappeared or remained during artery superselection. CONCLUSION: Transauricular cerebral angiography appears to be a feasible technique for brain or carotid intervention studies in rabbits. In addition, vertebral angiography using a transauricular approach is possible using the looping technique. Selection of carotid or vertebral arteries on each side was not difficult when the microcatheter and guide wire were looped within the cardiac chamber. The ear chosen for the initial puncture does not appear to be important.

Protein measurements in venous plasma, earlobe capillary plasma and in plasma stored on filter paper.

In this study, levels of inflammatory protein biomarkers in venous plasma, plasma derived from capillary blood from the earlobe, and capillary plasma stored as dried plasma spots (DPS) were compared. Samples from 12 male individuals were assessed with a panel of 92 inflammation-related proteins using multiplex proximity extension assay. Correlations between sample types varied greatly between analytes. A high correlation of ρ > 0.8 was observed between capillary plasma and DPS for 32 analytes. At this level of correlation, 13 analytes correlated between venous and capillary plasma and 5 analytes in the comparison of venous blood with DPS.

Significant Delay in the Detection of Desaturation between Finger Transmittance and Earlobe Reflectance Oximetry Probes during Fiberoptic Bronchoscopy: Analysis of 104 Cases.

PURPOSE: There is clinical significance to a delay in response time for detecting desaturation by pulse oximetry. Our aim in this study was to compare the response time of the reflectance and transmittance saturation probes during fiberoptic bronchoscopy (FOB) under monitored anesthesia care. METHODS: A prospective study included 104 patients scheduled for FOB. Patients were monitored with transmittance (finger) and reflectance (ear) oximetry probes. The response time was evaluated during desaturation and resaturation. We also acquired blood tests for arterial oxygen saturation to assess the agreement with the oximetry probes. RESULTS: Ninety patients had a desaturation episode during FOB and were included in the final analysis. Mean time difference between the reflectance ear probe (reference probe) and transmittance finger probe for the detection of desaturation (SpO2 = 90%) was + 36 s (CI 27.0-45.0, P < 0.001). The time difference between probes at end of desaturation episode (SpO2 = 95%) was + 31 s (CI 19.0-43.0; P < 0.001). A significant difference in response time was evident throughout the episode in all saturation values. The reflectance ear probe showed better agreement with arterial blood gases. The bias (and precision) for the earlobe and finger oximeters were of 0.24 (1.04) and 2.31 (3.37), respectively. CONCLUSION: The data displayed by a centrally located reflectance probe are more accurate and allows for earlier identification, treatment, and resolution of desaturation events. In light of these data and the added value of the reflectance probe ability to measure transcutaneous PCO2, we recommend monitoring bronchoscopy by a reflectance oximetry probe.

Experience of Replantation and Reconstruction in Total Scalp, Partial Forehead, and Ear Avulsions.

In this study, the authors present the challenges of replantation and management of 2 patients who have total scalp, partial forehead and ear avulsion. A 39-year-old male farmer was admitted to the emergency room with the total scalp, forehead, partial right eyebrow and upper eyelid amputation after his hair was catched by an agricultural machine. Anastomoses of 2 arteries and 2 veins in the occipital region were done first. Anastomosis of the right and left superficial temporal arteries and veins were performed end to end by turning the operating table without moving the head, followed by 1 vein anastomoses in the frontal region. Although anastomotic blood flow was observed by Doppler ultrasonography, there were changes suggesting necrosis in the frontal and both temporoparietal regions and later in the occipital region. Necrotic tissues began to be debrided tangentially from the 10th day. During serial debridement, it was observed that the necrotic tissue includes skin and connective tissue. At the end of the post-operative second month, surgical treatment was completed and 12 months after the surgery, the patient lives with the wig because of hair loss, but the patient is cosmetically satisfied. An 18-year-old female gatherer was admitted to the emergency room with the total scalp, both eyebrows, right upper eyelid and partial right ear avulsion, after her hair was catched by an agricultural machine. Totally, anastomoses of 4 arteries and 3 veins were performed. On the 10th post-operative day, necrotic skin findings appeared and serial debridement were performed and it was revealed that aponeurosis was fully intact. The defect areas of skin and connective tissue were repaired with STSG. Intact partial temporal scalp tissue was present. Scalp replantation should always be considered as the first choice, according to the similar tissue principle of tissue repair in plastic surgery if there is no contraindication. Additionally, performing multiple arterial and vein anastomoses, evaluating the patient position in the post-operative period for anastomosis and hemodynamic follow-up, avoiding aggressive debridement and early reconstruction may give the patient the chance of skin grafting which is the basis of the reconstructive ladder.

Real-time full-field optical angiography utilizing principal component analysis.

We develop a real-time full-field optical angiography method using principal component analysis (PCA). In our approach, an undersampled laser Doppler method is used to record the raw images. Considering the difference in the signal component contributions, PCA is used to separate the dynamic blood flow and static background signals. The principal advantage of the PCA method is that the choice of a high pixel number can aid in efficiently extracting the blood flow signal with finite frame raw images, which can greatly improve the temporal resolution. Our phantom experimental results validate our choice of the optimal frame number for reconstructing an angiographic image. A vascular occlusion test on a rabbit ear demonstrates that global and simultaneous hemodynamic processes of vessels can be monitored.

All-optical photoacoustic Doppler transverse blood flow imaging.

The method of measuring blood flow in photoacoustic microscopy usually relies on ultrasonic transducers in contact fashion, which is not favored in many applications, such as wound areas, burns, and anabrosis. Here we present a noncontact photoacoustic velocity measurement method to quantitatively map transverse blood flow based on the photoacoustic Doppler (PAD) bandwidth broadening method with an all-optical photoacoustic microscopy system. It is validated that the PAD bandwidth broadening is proportional to the transverse flow within a certain range. The transverse flow speed ranging from 0 to 5.5 mm/s, as well as sectional flow images, was obtained in the blood-mimicking flow phantoms. Furthermore, the blood flow image of the mouse ear demonstrates that the all-optical photoacoustic Doppler method can acquire the information of blood flow in vivo, which could significantly broaden the scope of applications for obtaining the blood flow velocity of the microvasculature in biomedicine.

Ultracompact high-resolution photoacoustic microscopy.

Optical resolution photoacoustic microscopy (ORPAM), benefiting from rich optical contrast, scalable acoustic resolution, and deep penetration depth, is of great importance for the fields of biology and medicine. However, limited by the size and performance of reported optical/acoustic scanners, existing portable/handheld ORPAMs are bulky and heavy, and suffer from low imaging quality/speed. Here, we present an ultracompact ORPAM probe, which is miniature and light, and has high imaging quality. The probe only weighs 20 grams and has an outer size of 22 mm×30 mm×13 mm, a high lateral resolution of 3.8 µm, and an effective imaging domain of 2 mm×2 mm. To show its advantages over existing ORPAMs, we apply this probe to image vasculatures of internal organs in a rat abdominal cavity and inspect the entire human oral cavity.

Fabrication of a multilayer tissue-mimicking phantom with tunable optical properties to simulate vascular oxygenation and perfusion for optical imaging technology.

Vast research has been carried out to fabricate tissue-mimicking phantoms, due to their convenient use and ease of storage, to assess and validate the performance of optical imaging devices. However, to the best of our knowledge, there has been little research on the use of multilayer tissue phantoms for optical imaging technology, although their structure is closer to that of real skin tissue. In this work, we design, fabricate, and characterize multilayer tissue-mimicking phantoms, with a morphological mouse ear blood vessel, that contain an epidermis, a dermis, and a hypodermis. Each tissue-mimicking phantom layer is characterized individually to match specific skin tissue layer characteristics. The thickness, optical properties (absorption coefficient and reduced scattering coefficient), oxygenation, and perfusion of skin are the most critical parameters for disease diagnosis and for some medical equipment. These phantoms can be used as calibration artifacts and help to evaluate optical imaging technologies.

Cross-correlation photothermal optical coherence tomography with high effective resolution.

We developed a cross-correlation photothermal optical coherence tomography (CC-PTOCT) system for photothermal imaging with high lateral and axial resolution. The CC-PTOCT system consists of a phase-sensitive OCT system, a modulated pumping laser, and a digital cross-correlator. The pumping laser was used to induce the photothermal effect in the sample, causing a slight phase modulation of the OCT signals. A spatial phase differentiation method was employed to reduce phase accumulation. The noise brought by the phase differentiation method and the strong background noise were suppressed efficiently by the cross-correlator, which was utilized to extract the photothermal signals from the modulated signals. Combining the cross-correlation technique with spatial phase differentiation can improve both lateral and axial resolution of the PTOCT imaging system. Clear photothermal images of blood capillaries of a mouse ear in vivo were successfully obtained with high lateral and axial resolution. The experimental results demonstrated that this system can enhance the effective transverse resolution, effective depth resolution, and contrast of the PTOCT image effectively, aiding the ongoing development of the accurate 3D functional imaging.

In vivo photoacoustic flow cytometry for early malaria diagnosis.

In vivo photoacoustic (PA) flow cytometry (PAFC) has already demonstrated a great potential for the diagnosis of deadly diseases through ultrasensitive detection of rare disease-associated circulating markers in whole blood volume. Here, we demonstrate the first application of this powerful technique for early diagnosis of malaria through label-free detection of malaria parasite-produced hemozoin in infected red blood cells (iRBCs) as high-contrast PA agent. The existing malaria tests using blood smears can detect the disease at 0.001-0.1% of parasitemia. On the contrary, linear PAFC showed a potential for noninvasive malaria diagnosis at an extremely low level of parasitemia of 0.0000001%, which is ∼10(3) times better than the existing tests. Multicolor time-of-flight PAFC with high-pulse repetition rate lasers at wavelengths of 532, 671, and 820 nm demonstrated rapid spectral and spatial identification and quantitative enumeration of individual iRBCs. Integration of PAFC with fluorescence flow cytometry (FFC) provided real-time simultaneous detection of single iRBCs and parasites expressing green fluorescence proteins, respectively. A combination of linear and nonlinear nanobubble-based multicolor PAFC showed capability to real-time control therapy efficiency by counting of iRBCs before, during, and after treatment. Our results suggest that high-sensitivity, high-resolution ultrafast PAFC-FFC platform represents a powerful research tool to provide the insight on malaria progression through dynamic study of parasite-cell interactions directly in bloodstream, whereas portable hand-worn PAFC device could be broadly used in humans for early malaria diagnosis. © 2016 International Society for Advancement of Cytometry.

Observation of vasculature alternation by intense pulsed light combined with physicochemical methods.

Intense pulsed light (IPL) with low energy insufficient to completely destroy a vasculature was applied to rabbit ears to investigate vasculature alteration. Glycerol was combined with IPL to enhance the transfer efficacy of IPL energy. Both trans-illumination and laser speckle contrast images were obtained and analyzed after treatment. The application of IPL and glycerol combination induced vasodilation and improvement in blood flow. Moreover, such phenomenon was maintained over time. IPL may be applied to treat blood circulatory diseases by inducing vasodilation and to improve blood flow.

The inhibitory effect of soybean and soybean isoflavone diets on 2,4-dinitrofluorobenzene-induced contact hypersensitivity in mice.

Murine contact hypersensitivity (CHS) is one of the most frequently used animal models of human allergic contact dermatitis. We investigated the inhibitory effects of soybean and soy isoflavone (SI) diets on 2,4-dinitrofluorobenzene- (DNFB) induced CHS in mice. The DNFB-induced ear swelling was inhibited in the soy- and SI-treated groups. Histopathological investigations revealed that oral feeding of soybean and SI attenuated ear tissue edema and reduced the number of Gr-1(+) cell infiltrations into ear tissues. DNA microarray analysis showed that the expression of Ccl24, Xcl1, Ifng, and Ccl17 in the ear tissues was lower in the soy-treated mice than in the positive controls. In addition, CCL24 mRNA and protein expression in the ear tissues were more highly suppressed in the soy- and SI-treated groups. These results suggest that soybean and SI consumption downregulated the gene and protein expression of CCL24, thereby affording protection against CHS in mice.

Investigation of histopathological effects after intra-arterial sugammadex injection in an experimental animal model.

PURPOSE: Intra-arterial injection of medications are related to mortality and morbidity. It was aimed to investigate the histopathological effects caused by two different intra-arterial doses of sugammadex, a new selective relaxant binding agent used to reverse the effects of steroidal systemic muscle relaxants used in surgeries carried out under general anesthesia. METHODS: A total of 15 rabbits were randomly divided into 3 groups. The control group was given 1 mL saline bolus into the central artery of the ear. The other two groups (Group Sgdx2 and group Sgdx16) were given 2 and 16 mg/kg sugammadex, respectively, intra-arterially. RESULTS: In control group, histopathological evaluation was in normal limits. The cross-sections belonging to Group Sgdx2 did not have any significant pathological change compared with control group (p > 0.05). Edema and single-cell necrosis were significantly higher in Group Sgdx16 when compared to control group (p < 0.05). In comparison to Group Sgdx2, single-cell necrosis was significantly higher in Group Sgdx16 (p < 0.05). CONCLUSION: Being broadly used in anesthesia, sugammadex should be administered with care because when given by intra-arterial route it may cause tissue damage and tissue loss and the patients' arteries should be carefully checked.

Simultaneous photoacoustic microscopy of microvascular anatomy, oxygen saturation, and blood flow.

Capitalizing on the optical absorption of hemoglobin, photoacoustic microscopy (PAM) is uniquely capable of anatomical and functional characterization of the intact microcirculation in vivo. However, PAM of the metabolic rate of oxygen (MRO2) at the microscopic level remains an unmet challenge, mainly due to the inability to simultaneously quantify microvascular diameter, oxygen saturation of hemoglobin (sO2), and blood flow at the same spatial scale. To fill this technical gap, we have developed a multi-parametric PAM platform. By analyzing both the sO2-encoded spectral dependence and the flow-induced temporal decorrelation of photoacoustic signals generated by the raster-scanned mouse ear vasculature, we demonstrated-for the first time-simultaneous wide-field PAM of all three parameters down to the capillary level in vivo.