Real-time line-field optical coherence tomography for cellular resolution imaging of biological tissue.

A real-time line-field optical coherence tomography (LF-OCT) system is demonstrated with image acquisition rates of up to 5000 B-frames or 2.5 million A-lines per second for 500 A-lines per B-frame. The system uses a high-speed low-cost camera to achieve continuous data transfer rates required for real-time imaging, allowing the evaluation of future applications in clinical or intraoperative environments. The light source is an 840 nm super-luminescent diode. Leveraging parallel computing with GPU and high speed CoaXPress data transfer interface, we were able to acquire, process, and display OCT data with low latency. The studied system uses anamorphic beam shaping in the detector arm, optimizing the field of view and sensitivity for imaging biological tissue at cellular resolution. The lateral and axial resolution measured in air were 1.7 µm and 6.3 µm, respectively. Experimental results demonstrate real-time inspection of the trabecular meshwork and Schlemm's canal on ex vivo corneoscleral wedges and real-time imaging of endothelial cells of human subjects in vivo.

Nanosensitive optical coherence tomography for detecting structural changes in stem cells.

Mesenchymal stromal cells (MSCs) are adult stem cells that have been widely investigated for their potential to regenerate damaged and diseased tissues. Multiple pre-clinical studies and clinical trials have demonstrated a therapeutic response following treatment with MSCs for various pathologies, including cardiovascular, neurological and orthopaedic diseases. The ability to functionally track cells following administration in vivo is pivotal to further elucidating the mechanism of action and safety profile of these cells. Effective monitoring of MSCs and MSC-derived microvesicles requires an imaging modality capable of providing both quantitative and qualitative readouts. Nanosensitive optical coherence tomography (nsOCT) is a recently developed technique that detects nanoscale structural changes within samples. In this study, we demonstrate for the first time, the capability of nsOCT to image MSC pellets following labelling with different concentrations of dual plasmonic gold nanostars. We show that the mean spatial period of MSC pellets increases following the labelling with increasing concentrations of nanostars. Additionally, with the help of extra time points and a more comprehensive analysis, we further improved the understanding of the MSC pellet chondrogenesis model. Despite the limited penetration depth (similar to conventional OCT), the nsOCT is highly sensitive in detecting structural alterations at the nanoscale, which may provide crucial functional information about cell therapies and their modes of action.

Corneal imaging with blue-light optical coherence microscopy.

Corneal imaging is important for the diagnostic and therapeutic evaluation of many eye diseases. Optical coherence tomography (OCT) is extensively used in ocular imaging due to its non-invasive and high-resolution volumetric imaging characteristics. Optical coherence microscopy (OCM) is a technical variation of OCT that can image the cornea with cellular resolution. Here, we demonstrate a blue-light OCM as a low-cost and easily reproducible system to visualize corneal cellular structures such as epithelial cells, endothelial cells, keratocytes, and collagen bundles within stromal lamellae. Our blue-light OCM system achieved an axial resolution of 12 µm in tissue over a 1.2 mm imaging depth, and a lateral resolution of 1.6 µm over a field of view of 750 µm × 750 µm.

Contrast agents for photoacoustic imaging: a review of stem cell tracking.

With the advent of stem cell therapy for spinal cord injuries, stroke, burns, macular degeneration, heart diseases, diabetes, rheumatoid arthritis and osteoarthritis; the need to track the survival, migration pathways, spatial destination and differentiation of transplanted stem cells in a clinical setting has gained increased relevance. Indeed, getting regulatory approval to use these therapies in the clinic depends on biodistribution studies. Although optoacoustic imaging (OAI) or photoacoustic imaging can detect functional information of cell activities in real-time, the selection and application of suitable contrast agents is essential to achieve optimal sensitivity and contrast for sensing at clinically relevant depths and can even provide information about molecular activity. This review explores OAI methodologies in conjunction with the specific application of exogenous contrast agents in comparison to other imaging modalities and describes the properties of exogenous contrast agents for quantitative and qualitative monitoring of stem cells. Specific characteristics such as biocompatibility, the absorption coefficient, and surface functionalization are compared and how the labelling efficiency translates to both short and long-term visualization of mesenchymal stem cells is explored. An overview of novel properties of recently developed optoacoustic contrast agents and their capability to detect disease and recovery progression in clinical settings is provided which includes newly developed exogenous contrast agents to monitor stem cells in real-time for multimodal sensing.

Accessing depth-resolved high spatial frequency content from the optical coherence tomography signal.

Optical coherence tomography (OCT) is a rapidly evolving technology with a broad range of applications, including biomedical imaging and diagnosis. Conventional intensity-based OCT provides depth-resolved imaging with a typical resolution and sensitivity to structural alterations of about 5-10 microns. It would be desirable for functional biological imaging to detect smaller features in tissues due to the nature of pathological processes. In this article, we perform the analysis of the spatial frequency content of the OCT signal based on scattering theory. We demonstrate that the OCT signal, even at limited spectral bandwidth, contains information about high spatial frequencies present in the object which relates to the small, sub-wavelength size structures. Experimental single frame imaging of phantoms with well-known sub-micron internal structures confirms the theory. Examples of visualization of the nanoscale structural changes within mesenchymal stem cells (MSC), which are invisible using conventional OCT, are also shown. Presented results provide a theoretical and experimental basis for the extraction of high spatial frequency information to substantially improve the sensitivity of OCT to structural alterations at clinically relevant depths.

Characterization of an amplified piezoelectric actuator for multiple reference optical coherence tomography: erratum.

This erratum is submitted to correct information regarding Fig. 8 of Appl. Opt.57, E142 (2018)APOPAI0003-693510.1364/AO.57.00E142.

Characterization of an amplified piezoelectric actuator for multiple-reference optical coherence tomography.

The characterization of an amplified piezoelectric actuator (APA) as a new axial scanning method for multiple-reference optical coherence tomography (MR-OCT) is described. MR-OCT is a compact optical imaging device based on a recirculating reference-arm-scanning optical delay using a partial mirror that can enhance the imaging depth range by more than 10 times the reference mirror's scanning amplitude. The scanning amplitude of the used APA was varied between 30 µm and 250 µm, depending on the scanning frequency of between 0.8 kHz and 1.2 kHz. A silver-coated miniature mirror was attached to the APA via ultraviolet-cured optical adhesive, and the light source was a super-luminescent diode with 1310 nm center wavelength and 56 nm bandwidth. The sensitivity was measured with and without the partial mirror in the reference delay line as a function of scan speed, frequency, and range, therefore providing results for MR-OCT and TD-OCT modes. It was found that the APA provides more than twice the mechanical scanning range compared to other opto-mechanic actuators, but results indicate degradation of signal-to-noise ratio and sensitivity at larger imaging depths. In conjunction with MR-OCT, the scan range of maximum 200 µm can be enhanced up to 1-1.5 mm by using a reduced amount of orders of reflections, which could be of interest to increase sensitivity in the future.

Simultaneous en-face imaging of multiple layers with multiple reference optical coherence tomography.

A technique based on multiple reference optical coherence tomography (MR-OCT) is proposed for simultaneous imaging at multiple depths. The technique has been validated by imaging a reference sample and a fingerprint in-vivo. The principle of scanning multiple selected layers is shown by imaging a partial fingerprint with 200×200×200 voxels of 3×3×0.5 mm size and obtaining an arbitrary amount of layers merely by digital processing. The spacing among the layers can be adjusted arbitrarily, and the SNR roll-off is shown for three different spacings. At a mirror scan frequency of 1 kHz and an A-line rate of 2 kHz, the acquisition time was 20 s for one volume. The results show the feasibility of the application of layer scanning MR-OCT that uses a partial mirror in the reference arm of the Michelson interferometer. The reduced scan range required for layer scanning allows even higher scan rates that are limited only by the voice coil design and the mass-spring system, e.g., mirror mass, spring constant, and damping.

Assessment of curing behavior of light-activated dental composites using intensity correlation based multiple reference optical coherence tomography.

BACKGROUND AND OBJECTIVES: Monitoring the curing kinetics of light-activated resin is a key area of research. These resins are used in restorative applications and particularly in dental applications. They can undergo volumetric shrinkage due to poor control of the depth dependent curing process, modulated by the intensity and duration of the curing light source. This often results in the formation of marginal gaps, causing pain and damage to the restoration site. In this study, we demonstrate the capabilities of a correlation method applied using a multiple references optical coherence tomography (MR-OCT) architecture to monitor the curing of the resin. STUDY DESIGN/MATERIALS AND METHODS: A MR-OCT system is used in this study to monitor the curing of the resin. The system operates at the center wavelength of 1310 nm with an A-scan rate of 1200 A-scans per second. The axial and lateral resolution of the system is ∼13 µm and ∼27 µm. The method to determine the intensity correlation between adjacent B-frames is based on the Pearson correlation coefficient for a region of interest. Calculating the correlation coefficient for multiple B-frames related to the first B-frame at regular spaced time points, shows for a noncured resin a reduction of the correlation coefficient over time due to Brownian motion. The time constant of the reduction of the correlation value is a measure for the progress of the polymerization during LED light irradiation of the resin. The proposed approach is potentially a low-cost, powerful and unique optical imaging modality for measuring the curing behavior of dental resin and other resins, coatings, and adhesives in medical and industrial applications. RESULTS: To demonstrate the proposed method to monitor the curing process, a light-activated resin composite from GRADIA DIRECT ANTERIOR (GC Corporation, Japan) is studied. The curing time of resin was measured and monitored as a function of depth. The correlation coefficient method is highly sensitive to Brownian motion. The process of curing results in a change in intensity as measured by the MR-OCT signal and hence can be monitored using this method. CONCLUSIONS: These results show that MR-OCT has the potential to measure the curing time and monitor the curing process as a function of depth. Moreover, MR-OCT as a product has potential to be compact, low-cost and to fit into a smartphone. Using such a device for monitoring the curing of the resin will be suitable for dentists in stationary and mobile clinical settings.

3D nondestructive testing system with an affordable multiple reference optical-delay-based optical coherence tomography.

Optical coherence tomography (OCT) is emerging as a powerful noncontact imaging technique, allowing high-quality cross-sectional imaging of scattering specimens nondestructively. However, the complexity and cost of current embodiments of an OCT system limit its use in various nondestructive testing (NDT) applications at resource-limited settings. In this paper, we demonstrate the feasibility of a novel low-cost OCT system for a range of nondestructive testing (NDT) applications. The proposed imaging system is based on an enhanced time-domain OCT system with a low cost and small form factor reference arm optical delay, called multiple reference OCT (MR-OCT), which uses a miniature voice coil actuator and a partial mirror for extending the axial scan range. The proposed approach is potentially a low-cost, compact, and unique optical imaging modality for a range of NDT applications in a low-resource setting. Using this method, we demonstrated the capability of MR-OCT to perform cross-sectional and volumetric imaging at 1200 A-scans per second.

Dermascope guided multiple reference optical coherence tomography.

In this paper, we report the feasibility of integrating a novel low cost optical coherence tomography (OCT) system with a dermascope for point-of-care applications. The proposed OCT system is based on an enhanced time-domain optical coherence tomographic system, called multiple reference OCT (MR-OCT), which uses a single miniature voice coil actuator and a partial mirror for extending the axial scan range. The system can simultaneously register both the superficial dermascope image and the depth-resolved OCT sub-surface information by an interactive beam steering method. A practitioner is able to obtain the depth resolved information of the point of interest by simply using the mouse cursor. The proposed approach of combining a dermascope with a low cost OCT provides a unique powerful optical imaging modality for a range of dermatological applications. Hand-held dermascopic OCT devices would also enable point of care and remote health monitoring.