Experimental Biology and Medicine

Fibrous tissues play important roles in many parts of the body. Their highly organized directional structure is essential in achieving their normal biomechanical and physiological functions. Disruption of the typical fiber organization in these tissues is often linked to pathological changes and disease progression. Tractography is a specialized imaging method that can reveal the detailed fiber architecture. Here, we review recent developments in high-resolution optical tractography using Jones matrix polarization-sensitive optical coherence tomography. We also illustrate the use of this new tractography technology for visualizing depth-resolved, three-dimensional fibrous structures and quantifying tissue damages in several major fibrous tissues.

Age-related macular degeneration (AMD) is a progressive retinal disease and becomes the leading cause of blindness. It is well established that early detection is the key to preservation of functional vision. However, it is very difficult to diagnose AMD in very early stages, before structural changes are evident. Consequently, investigating the biomechanical properties of the retina maybe essential for understanding its physiological function. In this study, we present a shear wave-based quantitative method for estimating the elasticity of the posterior eye using shaker-based optical coherence elastography. This technique has been developed and validated on both a homogeneous phantom and a healthy rabbit in vivo. The shear wave speed from the ganglion side to the photoreceptor side of the rabbit eye is 4.1 m/s, 4.9 m/s, and 6.7 m/s, respectively. In addition, the most stiff sclera region has an average shear wave speed of 9.1 m/s. The results demonstrate the feasibility of using this technique to quantify biomechanical properties of the posterior eye and its potential translation to the clinical study.

Noninvasive transcorneal electrical stimulation (TES) has emerged as a potential strategy to facilitate visual restoration and promote retinal cell survival for certain retinal and optic nerve diseases owing to its neuroprotective effects. However, the neurovascular responses of retinal neurons evoked by TES have not been completely determined. To investigate this issue, we utilized a custom-designed spectral-domain optical coherence tomography (SD-OCT) to record the retinal neural and vascular responses under TES in vivo simultaneously. Significant increases of both positive and negative intrinsic optical signal (IOS) changes were recorded in all three segmented retinal layers, which mainly related to neural activities. However, the changes of TES-induced retinal vascular responses, including blood velocity, cross-sectional area of vessel, and blood flow, were not significant. It suggests that TES mainly elicited neural responses in retina, while no significant vascular responses were evoked. Our results provide experimental evidence to the mechanism of retinal neurovascular coupling under TES. Additionally, the present study also suggests that SD-OCT could be utilized as a promoting method to explore neurovascular responses under retinal stimulation in clinical treatment and technology.

As a new optical coherence tomography (OCT) modality, OCT angiography (OCTA) provides a noninvasive method to detect microvascular distortions correlated with eye conditions. By providing unparalleled capability to differentiate individual plexus layers in the retina, OCTA has demonstrated its excellence in clinical management of diabetic retinopathy, glaucoma, sickle cell retinopathy, diabetic macular edema, and other eye diseases. Quantitative OCTA analysis of retinal and choroidal vasculatures is essential to standardize objective interpretations of clinical outcome. Quantitative features, including blood vessel tortuosity, blood vessel caliber, blood vessel density, vessel perimeter index, fovea avascular zone area, fovea avascular zone contour irregularity, vessel branching coefficient, vessel branching angle, branching width ratio, and choroidal vascular analysis have been established for objective OCTA assessment. Moreover, differential artery–vein analysis has been recently demonstrated to improve OCTA performance for objective detection and classification of eye diseases. In this review, technical rationales and clinical applications of these quantitative OCTA features are summarized, and future prospects for using these quantitative OCTA features for artificial intelligence classification of eye conditions are discussed.

To pave the road toward clinical application of photoacoustic imaging in prostate cancer (PCa) diagnosis, we studied the technical feasibility and performance of transrectal photoacoustic (PA) imaging in mapping the indocyanine green (ICG) contrast agent, which is approved by FDA, in entire prostates by using light illumination via the urethral track. Experiments were conducted on a clinically relevant ex vivo model involving whole human prostates harvested from radical prostatectomy. The light source placed in the urethral track was an array of light emitting diodes (LEDs), illuminating the prostate with a delivered light power on the urethral wall within the safety limit. A dual-modality imaging system acquired PA and ultrasound (US) images simultaneously in the same way as in transrectal ultrasound (TRUS), with the US imaging presenting the tissue structure and PA imaging detecting the ICG solution. The imaging results demonstrated that tubes containing ICG solution at different concentrations can be detected at different positions in the prostate within a 2 cm range around from the urethral wall. Considering the sizes of regular human prostates, the proposed transurethral illumination in combination with transrectal US detection can facilitate PA molecular imaging over the entire prostate in a non-invasive manner, which makes it possible to further improve the PCa diagnosing efficiency with better molecular sensitivity and resulted better biopsy accuracy and much reduced pain for patients.

Photoacoustic imaging is a non-invasive and non-ionizing biomedical technique that has been investigated widely for various clinical applications. By taking the advantages of conventional ultrasound imaging, hand-held operation with a linear array transducer should be favorable for successful clinical translation of photoacoustic imaging. In this paper, we present new key updates contributed to the previously developed real-time clinical photoacoustic and ultrasound imaging system for improving the clinical usability of the system. We developed a seamless image optimization platform, designed a real-time parameter control software with a user-friendly graphical user interface, performed Monte Carlo simulation of the optical fluence in the imaging plane, and optimized the geometry of the imaging probe. The updated system allows optimizing of all imaging parameters while continuously acquiring the photoacoustic and ultrasound images in real-time. The updated system has great potential to be used in a variety of clinical applications such as assessing the malignancy of thyroid cancer, breast cancer, and melanoma.

Photoacoustic imaging has demonstrated its potential for diagnosis over the last few decades. In recent years, its unique imaging capabilities, such as detecting structural, functional and molecular information in deep regions with optical contrast and ultrasound resolution, have opened up many opportunities for photoacoustic imaging to be used during image-guided interventions. Numerous studies have investigated the capability of photoacoustic imaging to guide various interventions such as drug delivery, therapies, surgeries, and biopsies. These studies have demonstrated that photoacoustic imaging can guide these interventions effectively and non-invasively in real-time. In this minireview, we will elucidate the potential of photoacoustic imaging in guiding active and passive drug deliveries, photothermal therapy, and other surgeries and therapies using endogenous and exogenous contrast agents including organic, inorganic, and hybrid nanoparticles, as well as needle-based biopsy procedures. The advantages of photoacoustic imaging in guided interventions will be discussed. It will, therefore, show that photoacoustic imaging has great potential in real-time interventions due to its advantages over current imaging modalities like computed tomography, magnetic resonance imaging, and ultrasound imaging.

We have developed a multimodal imaging system, which integrated optical resolution photoacoustic microscopy, optical coherence tomography, optical coherence tomography angiography, and confocal fluorescence microscopy in one platform. The system is able to image complementary features of a biological sample by combining different contrast mechanisms. We achieved fast imaging and large field of view by combining optical scanning with mechanical scanning, similar to our previous publication. We have demonstrated the capability of the multimodal imaging system by imaging a mouse ear in vivo.

Chondrocyte viability is a crucial factor for evaluating cartilage health. Most prevalent cell viability assays rely on dyes and are not applicable for in vivo or longitudinal studies. Here we demonstrated that the two-photon excited autofluorescence and second harmonic generation microscopy provided high-resolution imaging of cartilage tissue and distinguished live/dead chondrocytes by visual assessment. Furthermore, the normalized autofluorescence ratio was proposed as a quantitative indicator to determine chondrocyte viability. Based on the indicator, a curve fitting and simulated receiver operating characteristic method was proposed to identify the live/dead cell populations as well as the indicator threshold without dye labeling. Thus, it established the label-free imaging method for chondrocyte viability assay in cartilage tissue.

Transient intrinsic optical signal (IOS) has been observed in stimulus-evoked retinal photoreceptors. This study is to compare IOS changes in wild-type and retinal degeneration 10 (rd10) mouse retinas, to evaluate the effect of cyclic guanosine monophosphate phosphodiesterase on photoreceptor-IOS. Time-lapse near-infrared light microscopy was employed to monitor the spatiotemporal dynamics of the IOS responses in freshly isolated retinas activated by visible light stimulation. Comparative IOS recordings were conducted at postnatal days 14 (P14) and P16. At P14, intrinsic optical signal magnitudes and spatiotemporal dynamics in wild-type and rd10 retinas were similar, indicating that the phosphodiesterase deficiency in rd10 did not affect the formation of photoreceptor-IOS. At P16, IOS magnitude in rd10 significantly decreased compared to that in wild-type, suggesting the IOS sensitivity to the photoreceptor degeneration in rd10. Our experimental results and theoretical analysis indicate that early disc-based stages of the phototransduction cascade before the activation of phosphodiesterase may contribute to the formation of the photoreceptor-IOS responses; and the IOS can be a sensitive biomarker for objective assessment of retinal function.

Optical mapping has become a widely used and important method in cardiac electrophysiology. The method typically uses voltage-sensitive fluorescent dyes and high-speed cameras to image propagation of electrical waves. However, signals are highly susceptible to artifact caused by motion of the target organ. Consequently, cardiac optical mapping is traditionally performed in isolated, perfused organs whose contraction has been pharmacologically arrested. This has prevented optical mapping from being used to study interactions between electrical and mechanical motion. However, recently, a number of groups have developed methods to implement cardiac optical mapping in the presence of motion. These methods employ two basic strategies: (1) compensate for motion by measuring it or (2) ratiometry. In ratiometry, two signals are recorded from each site. The signals have differing sensitivity to membrane potential, but common motion artifact, which can be cancelled by taking the ratio of the two signals. Some methods use both of these strategies. Methods that measure motion have the additional advantage that this information can be used to quantify the organ’s mechanical function. Doing so enables combined “electromechanical mapping,” which allows optical study of electromechanical interactions. By allowing recording in the presence of motion, the new methods open the door to optical recording in in-vivo preparations. In addition, it is possible to implement electromechanical optical mapping techniques in organ systems other than the heart. For example, it was recently shown that optical mapping of slow wave propagation in the swine stomach is feasible. Such studies have the potential to uncover new information on the role of dysrhythmic slow wave propagation in gastric motility disorders.

Mutations in ACTA2, encoding smooth muscle α-actin, are a frequent cause of heritable thoracic aortic aneurysm and dissections. These mutations are associated with impaired vascular smooth muscle cell function, which leads to decreased ability of the cell to sense matrix-mediated mechanical stimuli. This study investigates how loss of smooth muscle α-actin affects cytoskeletal tension development and cell adhesion using smooth muscle cells explanted from aorta of mice lacking smooth muscle α-actin. We tested the hypothesis that reduced vascular smooth muscle contractility due to a loss of smooth muscle α-actin decreases cellular mechanosensing by dysregulating cell adhesion to the matrix. Assessment of functional mechanical properties of the aorta by stress relaxation measurements in thoracic aortic rings suggested two functional regimes for Acta2^−/− mice. Lower stress relaxation was recorded in aortic rings from Acta2^−/− mice at tensions below 10 mN compared with wild type, likely driven by cytoskeletal-dependent contractility. However, no differences were recorded between the two groups above the 10 mN threshold, since at higher tension the matrix-dependent contractility may be predominant. In addition, our results showed that at any given level of stretch, transmural pressure is lower in aortic rings from Acta2^−/− mice than wild type mice. In addition, a three-dimensional collagen matrix contractility assay showed that collagen pellets containing Acta2^−/− smooth muscle cells contracted less than the pellets containing the wild type cells. Moreover, second harmonic generation non-linear microscopy revealed that Acta2^−/− cells locally remodeled the collagen matrix fibers to a lesser extent than wild type cells. Quantification of protein fluorescence measurements in cells also showed that in absence of smooth muscle α-actin, there is a compensatory increase in smooth muscle γ-actin. Moreover, specific integrin recruitment at cell–matrix adhesions was reduced in Acta2^−/− cells. Thus, our findings suggest that Acta2^−/− cells are unable to generate external forces to remodel the matrix due to reduced contractility and interaction with the matrix.