Thermally tuned VCSEL at 850†nm as a low-cost alternative source for full-eye SS-OCT.

Swept-source optical coherence tomography (SS-OCT) demonstrates superior performance in comparison to spectral domain OCT with regard to depth ranging. The main driver of cost for SS-OCT systems is, however, the price of the source. Here we show a low-cost alternative swept source that uses a thermally tuned vertical-cavity surface-emitting laser (VCSEL) at 850 nm. Its center wavelength can be tuned by adjusting the operating temperature through modulation of the injection current. At 2 kHz sweep rate, the depth range of the system was 5 cm, with a sensitivity roll-off of under -3 dB across this range. The system achieved a sensitivity of 97 dB with a sample beam power of 0.3 mW and an axial resolution of 50 µm in air. To demonstrate the system performance in vivo, an eye of a healthy volunteer was measured, and full-eye scans were acquired at 25 and 50 kHz from the cornea to the retina. Based on our results, we believe that this technology can be used as a cost-effective alternative OCT for point-of-care diagnostics.

Depixelation of coherent fiber bundle imaging by fiber-core-targeted scanning.

A novel fast proximal scanning method, to the best of our knowledge, termed fiber-core-targeted scanning (FCTS), is proposed for illuminating individual fiber cores sequentially to remove the pixelation effect in fiber bundle (FB) imaging. FCTS is based on a galvanometer scanning system. Through a dynamic control of the scan trajectory and speed using the prior knowledge of fiber core positions, FCTS experimentally verifies a precise sequential delivery of laser pulses into fiber cores at a maximal speed of 45,000 cores per second. By applying FCTS on a FB-based photoacoustic forward-imaging probe, the results demonstrate that FCTS eliminates the pixelation effect and improves the imaging quality.

A Quantitative Model for Optical Coherence Tomography.

Optical coherence tomography (OCT) is a widely used imaging technique in the micrometer regime, which gained accelerating interest in medical imaging in the last twenty years. In up-to-date OCT literature, certain simplifying assumptions are made for the reconstructions, but for many applications, a more realistic description of the OCT imaging process is of interest. In mathematical models, for example, the incident angle of light onto the sample is usually neglected or a plane wave description for the light-sample interaction in OCT is used, which ignores almost completely the occurring effects within an OCT measurement process. In this article, we make a first step to a quantitative model by considering the measured intensity as a combination of back-scattered Gaussian beams affected by the system. In contrast to the standard plane wave simplification, the presented model includes system relevant parameters, such as the position of the focus and the spot size of the incident laser beam, which allow a precise prediction of the OCT data. The accuracy of the proposed model-after calibration of all necessary system parameters-is illustrated by simulations and validated by a comparison with experimental data obtained from a 1300 nm swept-source OCT system.

Three-dimensional assessment of para- and perifoveal photoreceptor densities and the impact of meridians and age in healthy eyes with adaptive-optics optical coherence tomography (AO-OCT).

An adaptive optics optical coherence tomography (AO-OCT) system is used to assess sixty healthy eyes of thirty subjects (age 22 to 75) to evaluate how the outer retinal layers, foveal eccentricity and age effect the mean cone density. The cone mosaics of different retinal planes (the cone inner segment outer segment junction (IS/OS), the cone outer segment combined with the IS/OS (ISOS+), the cone outer segment tips (COST), and the full en-face plane (FEF)) at four main meridians (superior, nasal, inferior, temporal) and para- and perifoveal eccentricities (ecc 2.5° and 6.5°) were analyzed quantitatively. The mean overall cone density was 19,892/mm2 at ecc 2.5° and 13,323/mm2 at ecc 6.5°. A significant impact on cone density was found for eccentricity (up to 6,700/mm2 between ecc 2.5° and 6.5°), meridian (up to 3,700/mm2 between nasal and superior meridian) and layer (up to 1,400/mm2 between FEF and IS/OS). Age showed only a weak negative effect. These factors as well as inter-individual variability have to be taken into account when comparing cone density measurements between healthy and pathologically changed eyes, as their combined effect on density can easily exceed several thousand cones per mm2 even in parafoveal regions.

Multi-channel swept source optical coherence tomography concept based on photonic integrated circuits.

In this paper, we present a novel concept for a multi-channel swept source optical coherence tomography (OCT) system based on photonic integrated circuits (PICs). At the core of this concept is a low-loss polarization dependent path routing approach allowing for lower excess loss compared to previously shown PIC-based OCT systems, facilitating a parallelization of measurement units. As a proof of concept for the low-loss path routing, a silicon nitride PIC-based single-channel swept source OCT system operating at 840 nm was implemented and used to acquire in-vivo tomograms of a human retina. The fabrication of the PIC was done via CMOS-compatible plasma-enhanced chemical vapor deposition to allow future monolithic co-integration with photodiodes and read-out electronics. A performance analysis using the results of the implemented photonic building blocks shows a potential tenfold increase of the acquisition speed for a multi-channel system compared to an ideal lossless single-channel system with the same signal-to-noise ratio.

Morpho-molecular ex vivo detection and grading of non-muscle-invasive bladder cancer using forward imaging probe based multimodal optical coherence tomography and Raman spectroscopy.

Non-muscle-invasive bladder cancer affects millions of people worldwide, resulting in significant discomfort to the patient and potential death. Today, cystoscopy is the gold standard for bladder cancer assessment, using white light endoscopy to detect tumor suspected lesion areas, followed by resection of these areas and subsequent histopathological evaluation. Not only does the pathological examination take days, but due to the invasive nature, the performed biopsy can result in significant harm to the patient. Nowadays, optical modalities, such as optical coherence tomography (OCT) and Raman spectroscopy (RS), have proven to detect cancer in real time and can provide more detailed clinical information of a lesion, e.g. its penetration depth (stage) and the differentiation of the cells (grade). In this paper, we present an ex vivo study performed with a combined piezoelectric tube-based OCT-probe and fiber optic RS-probe imaging system that allows large field-of-view imaging of bladder biopsies, using both modalities and co-registered visualization, detection and grading of cancerous bladder lesions. In the present study, 119 examined biopsies were characterized, showing that fiber-optic based OCT provides a sensitivity of 78% and a specificity of 69% for the detection of non-muscle-invasive bladder cancer, while RS, on the other hand, provides a sensitivity of 81% and a specificity of 61% for the grading of low- and high-grade tissues. Moreover, the study shows that a piezoelectric tube-based OCT probe can have significant endurance, suitable for future long-lasting in vivo applications. These results also indicate that combined OCT and RS fiber probe-based characterization offers an exciting possibility for label-free and morpho-chemical optical biopsies for bladder cancer diagnostics.

Ultrashort pulse Kagome hollow-core photonic crystal fiber delivery for nonlinear optical imaging.

We report ultrashort pulse delivery through a hypocycloid-core inhibited-coupling Kagome hollow-core photonic crystal fiber (HC-PCF). Undistorted 10 fs and 6.6 nJ pulses were launched through 1 m long fiber without fiber dispersion pre-compensation and 80% efficiency. The performance of this technology for biomedical imaging is demonstrated on a biological sample by incorporating the fiber into a two-photon excited fluorescence (TPEF) laser scanning microscope (LSM) achieving a pulse width of 15 fs at the sample location. To the best of our knowledge, this is the first report on undistorted TPEF imaging in a LSM with 15 fs pulses delivered through a 1 m long Kagome HC-PCF with high throughput.

Optical coherence tomography angiography and photoacoustic imaging in dermatology.

Optical coherence tomography angiography (OCTA) is a relatively novel functional extension of the widely accepted ophthalmic imaging tool named optical coherence tomography (OCT). Since OCTA's debut in ophthalmology, researchers have also been trying to expand its translational application in dermatology. The ability of OCTA to resolve microvasculature has shown promising results in imaging skin diseases. Meanwhile, photoacoustic imaging (PAI), which uses laser pulse induced ultrasound waves as the signal, has been studied to differentiate human skin layers and to help in skin disease diagnosis. This perspective article gives a short review of OCTA and PAI in the field of photodermatology. After an introduction to the principles of OCTA and PAI, we describe the most updated results of skin disease imaging using these two optical imaging modalities. We also place emphasis on dual modality imaging combining OCTA and photoacoustic tomography (PAT) for dermatological applications. In the end, the challenges and prospects of these two imaging modalities in dermatology are discussed.

Depth resolved label-free multimodal optical imaging platform to study morpho-molecular composition of tissue.

Multimodal imaging platforms offer a vast array of tissue information in a single image acquisition by combining complementary imaging techniques. By merging different systems, better tissue characterization can be achieved than is possible by the constituent imaging modalities alone. The combination of optical coherence tomography (OCT) with non-linear optical imaging (NLOI) techniques such as two-photon excited fluorescence (TPEF), second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) provides access to detailed information of tissue structure and molecular composition in a fast, label-free and non-invasive manner. We introduce a multimodal label-free approach for morpho-molecular imaging and spectroscopy and validate the system in mouse skin demonstrating the potential of the system for colocalized acquisition of OCT and NLOI signals.

THREE-DIMENSIONAL ANALYSIS OF RETINAL MICROANEURYSMS WITH ADAPTIVE OPTICS OPTICAL COHERENCE TOMOGRAPHY.

PURPOSE: To characterize retinal microaneurysms (MAs) in patients with diabetes using adaptive optics optical coherence tomography (AOOCT) and compare details found in AOOCT with those found in commercially available retinal imaging techniques. METHODS: Patients with diabetes and MA in the macular area were included in this pilot study. The area of interest, identified in standard fluorescein angiography, was imaged using an AO fundus camera and AOOCT. Microaneurysms were characterized in AOOCT (visibility, reflectivity, feeding/draining vessels, and intraretinal location) and compared with findings in AO fundus camera, OCT angiography, and fluorescein angiography. RESULTS: Fifty-three MAs were imaged in 15 eyes of 10 patients. Feeding and/or draining vessels from both capillary plexus could be identified in 34 MAs in AOOCT images. Of 45 MAs imaged with OCT angiography, 18 (40%) were visible in the superior plexus, 12 (27%) in the deep capillary plexus, and 15 MAs (33%) could not be identified at all. Intraluminal hyperreflectivity, commonly seen in AO fundus camera, corresponded only in 8 of 27 cases (30%) to intraluminal densities seen in AOOCT. CONCLUSION: Adaptive optics OCT imaging revealed that MAs located in the inner nuclear layer were connected to the intermediate and/or deep capillary plexus. Intraluminal hyperreflectivity seen on AO fundus camera images originated from a strong reflection from the vessel wall and only in a third of the cases from intraluminal clots. Currently, AOOCT is the most expedient in vivo imaging method to capture morphologic details of retinal microvasculature in 3D and in the context of the surrounding retinal anatomy.

Line Scan Raman Microspectroscopy for Label-Free Diagnosis of Human Pituitary Biopsies.

Pituitary adenomas are neoplasia of the anterior pituitary gland and can be subdivided into hormone-producing tumors (lactotroph, corticotroph, gonadotroph, somatotroph, thyreotroph or plurihormonal) and hormone-inactive tumors (silent or null cell adenomas) based on their hormonal status. We therefore developed a line scan Raman microspectroscopy (LSRM) system to detect, discriminate and hyperspectrally visualize pituitary gland from pituitary adenomas based on molecular differences. By applying principal component analysis followed by a k-nearest neighbor algorithm, specific hormone states were identified and a clear discrimination between pituitary gland and various adenoma subtypes was achieved. The classifier yielded an accuracy of 95% for gland tissue and 84-99% for adenoma subtypes. With an overall accuracy of 92%, our LSRM system has proven its potential to differentiate pituitary gland from pituitary adenomas. LSRM images based on the presence of specific Raman bands were created, and such images provided additional insight into the spatial distribution of particular molecular compounds. Pathological states could be molecularly differentiated and characterized with texture analysis evaluating Grey Level Cooccurrence Matrices for each LSRM image, as well as correlation coefficients between LSRM images.

Dual modality reflection mode optical coherence and photoacoustic microscopy using an akinetic sensor: publisher's note.

This publisher's note corrects an error in the funding section in Opt. Lett.42, 4319 (2017)OPLEDP0146-959210.1364/OL.42.004319.

Dual modality reflection mode optical coherence and photoacoustic microscopy using an akinetic sensor.

This Letter presents a novel dual modality reflection mode optical coherence and photoacoustic microscopy (OC-PAM) system. The optical coherence microscopy modality features a broadband source to accomplish 5 µm axial resolution. The photoacoustic microscopy modality uses a rigid akinetic Fabry-Perot etalon encapsulated in an optically transparent medium, which forms a 2 mm×11 mm translucent imaging window, permitting reflection mode dual modality imaging. After characterization, the OC-PAM system was applied to image zebrafish larvae in vivo, demonstrating its capability in biomedical imaging with complementary optical scattering and absorption contrasts by revealing morphology in the fish larvae.

Mapping diurnal changes in choroidal, Haller's and Sattler's layer thickness using 3-dimensional 1060-nm optical coherence tomography.

PURPOSE: To test the significance of diurnal changes in choroidal, Haller's and Sattler's layer thickness in healthy subjects using spatial analysis of three-dimensional (3D) 1060-nm optical coherence tomography (OCT) scans. METHODS: Automatically generated choroidal, Haller's and Sattler's layer thickness maps were statistically analyzed for 19 healthy subjects at two time points (8 a.m. and 6 p.m.) that represent the currently proposed ChT peak and nadir. All subjects were imaged by high-speed 1060-nm OCT over a 36° × 36° field of view. Spatial distribution of layer thickness was analyzed using the Early Treatment Diabetic Retinopathy Study (ETDRS) grid. RESULTS: The choroid was significantly thicker at 8 a.m. than at 6 p.m. (p < 0,0125, paired t-test, Bonferroni correction). Diurnal variation of mean choroidal thickness (ChT) for all ETDRS subfields was 12 µm. Haller's layer thickness showed no significant diurnal variation (P > 0.0125), but Sattler's layer was thicker in the morning than in late afternoon (P < 0.0125). CONCLUSIONS: Our measurements indicate that diurnal ChT variation may exist, but is less relevant than previously proposed by studies using single location imaging. Sattler's layer shows diurnal variation in line with ChT.

Single-pulse CARS based multimodal nonlinear optical microscope for bioimaging.

Noninvasive label-free imaging of biological systems raises demand not only for high-speed three-dimensional prescreening of morphology over a wide-field of view but also it seeks to extract the microscopic functional and molecular details within. Capitalizing on the unique advantages brought out by different nonlinear optical effects, a multimodal nonlinear optical microscope can be a powerful tool for bioimaging. Bringing together the intensity-dependent contrast mechanisms via second harmonic generation, third harmonic generation and four-wave mixing for structural-sensitive imaging, and single-beam/single-pulse coherent anti-Stokes Raman scattering technique for chemical sensitive imaging in the finger-print region, we have developed a simple and nearly alignment-free multimodal nonlinear optical microscope that is based on a single wide-band Ti:Sapphire femtosecond pulse laser source. Successful imaging tests have been realized on two exemplary biological samples, a canine femur bone and collagen fibrils harvested from a rat tail. Since the ultra-broad band-width femtosecond laser is a suitable source for performing high-resolution optical coherence tomography, a wide-field optical coherence tomography arm can be easily incorporated into the presented multimodal microscope making it a versatile optical imaging tool for noninvasive label-free bioimaging.

Numerical focusing methods for full field OCT: a comparison based on a common signal model.

In this paper a theoretical model of the full field swept source (FF SS) OCT signal is presented based on the angular spectrum wave propagation approach which accounts for the defocus error with imaging depth. It is shown that using the same theoretical model of the signal, numerical defocus correction methods based on a simple forward model (FM) and inverse scattering (IS), the latter being similar to interferometric synthetic aperture microscopy (ISAM), can be derived. Both FM and IS are compared quantitatively with sub-aperture based digital adaptive optics (DAO). FM has the least numerical complexity, and is the fastest in terms of computational speed among the three. SNR improvement of more than 10 dB is shown for all the three methods over a sample depth of 1.5 mm. For a sample with non-uniform refractive index with depth, FM and IS both improved the depth of focus (DOF) by a factor of 7x for an imaging NA of 0.1. DAO performs the best in case of non-uniform refractive index with respect to DOF improvement by 11x.

Line-field parallel swept source interferometric imaging at up to 1 MHz.

We present a novel medical imaging modality based on optical coherence tomography (OCT) that enables in vivo 3D tomography at acquisition rates up to 1 MHz. Line field parallel swept source interferometric imaging (LPSI) combines line-field swept source OCT with modulation of the interferometric signal in spatial direction for full range imaging. This method enables high speed imaging with cost-effective and commercially available technology. We explain the realization of the LPSI setup, acquisition, and postprocessing and finally demonstrate 3D in vivo imaging of human nail fold. To the best of our knowledge, sensitivity and depth penetration are competitive with respective point scanning OCT methods at a comparable wavelength. Measured maximum sensitivity is 98.5 dB for 100 kHz and 90 dB for 1 MHz. Together with the significantly relaxed technological requirements regarding detection and swept source technology, LPSI might be a promising concept for future diagnostic OCT imaging.

Hybrid single-source online Fourier transform coherent anti-Stokes Raman scattering/optical coherence tomography.

We demonstrate a multimodal optical coherence tomography (OCT) and online Fourier transform coherent anti-Stokes Raman scattering (FTCARS) platform using a single sub-12 femtosecond (fs) Ti:sapphire laser enabling simultaneous extraction of structural and chemical ("morphomolecular") information of biological samples. Spectral domain OCT prescreens the specimen providing a fast ultrahigh (4×12 µm axial and transverse) resolution wide field morphologic overview. Additional complementary intrinsic molecular information is obtained by zooming into regions of interest for fast label-free chemical mapping with online FTCARS spectroscopy. Background-free CARS is based on a Michelson interferometer in combination with a highly linear piezo stage, which allows for quick point-to-point extraction of CARS spectra in the fingerprint region in less than 125 ms with a resolution better than 4 cm(-1) without the need for averaging. OCT morphology and CARS spectral maps indicating phosphate and carbonate bond vibrations from human bone samples are extracted to demonstrate the performance of this hybrid imaging platform.

Ultrasound-induced reorientation for multi-angle optical coherence tomography.

Organoid and spheroid technology provide valuable insights into developmental biology and oncology. Optical coherence tomography (OCT) is a label-free technique that has emerged as an excellent tool for monitoring the structure and function of these samples. However, mature organoids are often too opaque for OCT. Access to multi-angle views is highly desirable to overcome this limitation, preferably with non-contact sample handling. To fulfil these requirements, we present an ultrasound-induced reorientation method for multi-angle-OCT, which employs a 3D-printed acoustic trap inserted into an OCT imaging system, to levitate and reorient zebrafish larvae and tumor spheroids in a controlled and reproducible manner. A model-based algorithm was developed for the physically consistent fusion of multi-angle data from a priori unknown angles. We demonstrate enhanced penetration depth in the joint 3D-recovery of reflectivity, attenuation, refractive index, and position registration for zebrafish larvae, creating an enabling tool for future applications in volumetric imaging.

Visualization of Cataract Surgery Steps With 4D Microscope-Integrated Swept-Source Optical Coherence Tomography in Ex Vivo Porcine Eyes.

Purpose: To investigate the visualization capabilities of high-speed swept-source optical coherence tomography (SS-OCT) in cataract surgery. Methods: Cataract surgery was simulated in wet labs with ex vivo porcine eyes. Each phase of the surgery was visualized with a novel surgical microscope-integrated SS-OCT with a variable imaging speed of over 1 million A-scans per second. It was designed to provide four-dimensional (4D) live-volumetric videos, live B-scans, and volume capture scans. Results: Four-dimensional videos, B-scans, and volume capture scans of corneal incision, ophthalmic viscosurgical device injection, capsulorrhexis, phacoemulsification, intraocular lens (IOL) injection, and position of unfolded IOL in the capsular bag were recorded. The flexibility of the SS-OCT system allowed us to tailor the scanning parameters to meet the specific demands of dynamic surgical steps and static pauses. The entire length of the eye was recorded in a single scan, and unfolding of the IOL was visualized dynamically. Conclusions: The presented novel visualization method for fast ophthalmic surgical microscope-integrated intraoperative OCT imaging in cataract surgery allowed the visualization of all major steps of the procedure by achieving large imaging depths covering the entire eye and high acquisition speeds enabling live volumetric 4D-OCT imaging. This promising technology may become an integral part of routine and advanced robotic-assisted cataract surgery in the future. Translational Relevance: We demonstrate the visualization capabilities of a cutting edge swept-source OCT system integrated into an ophthalmic surgical microscope during cataract surgery.