Optical coherence tomography-guided Brillouin microscopy highlights regional tissue stiffness differences during anterior neural tube closure in the Mthfd1l murine mutant.

Neurulation is a highly synchronized biomechanical process leading to the formation of the brain and spinal cord, and its failure leads to neural tube defects (NTDs). Although we are rapidly learning the genetic mechanisms underlying NTDs, the biomechanical aspects are largely unknown. To understand the correlation between NTDs and tissue stiffness during neural tube closure (NTC), we imaged an NTD murine model using optical coherence tomography (OCT), Brillouin microscopy and confocal fluorescence microscopy. Here, we associate structural information from OCT with local stiffness from the Brillouin signal of embryos undergoing neurulation. The stiffness of neuroepithelial tissues in Mthfd1l null embryos was significantly lower than that of wild-type embryos. Additionally, exogenous formate supplementation improved tissue stiffness and gross embryonic morphology in nullizygous and heterozygous embryos. Our results demonstrate the significance of proper tissue stiffness in normal NTC and pave the way for future studies on the mechanobiology of normal and abnormal embryonic development.

Red light instruments for myopia exceed safety limits.

PURPOSE: Low-level red light (LLRL) therapy has recently emerged as a myopia treatment in children, with several studies reporting significant reduction in axial elongation and myopia progression. The goal of this study was to characterise the output and determine the thermal and photochemical maximum permissible exposure (MPE) of LLRL devices for myopia control. METHODS: Two LLRL devices, a Sky-n1201a and a Future Vision, were examined. Optical power measurements were made using an integrating sphere radiometer through a 7-mm diameter aperture, in accordance with ANSI Z136.1-2014, sections 3.2.3-3.2.4. Retinal spot sizes of the devices were obtained using a model eye and high-resolution beam profiler. Corneal irradiance, retinal irradiance and MPE were calculated for an eye positioned at the oculars of each device. RESULTS: Both devices were confirmed to be Class 1 laser products. Findings showed that the Sky-n1201a delivers laser light as a point source with a 654-nm wavelength, 0.2 mW power (Ø 7 mm aperture, 10-cm distance), 1.17 mW/cm2 corneal irradiance and 7.2 W/cm2 retinal irradiance (Ø 2 mm pupil). The MPE for photochemical damage is 0.55-7.0 s for 2-7 mm pupils and for thermal damage is 0.41-10 s for 4.25-7 mm pupils. Future Vision delivers the laser as an extended source subtending 0.75 × 0.325°. It has a 652-nm wavelength, 0.06 mW power (Ø 7 mm aperture, 10 cm distance), 0.624 mW/cm2 corneal irradiance and 0.08 W/cm2 retinal irradiance (Ø 2 mm pupil). MPE for photochemical damage is 50-625 s for 2-7 mm pupils. DISCUSSION: For both of the LLRL devices evaluated here, 3 min of continuous viewing approached or surpassed the MPE, putting the retina at risk of photochemical and thermal damage. Clinicians should be cautious with the use of LLRL therapy for myopia in children until safety standards can be confirmed.

Reverberant optical coherence elastography using multifocal acoustic radiation force.

In this study, we introduce a multifocal acoustic radiation force source that combines an ultrasound transducer and a 3D-printed acoustic lens for application in reverberant optical coherence elastography (Rev-OCE). An array of plano-concave acoustic lenses, each with an 11.8 mm aperture diameter, were used to spatially distribute the acoustic energy generated by a 1 MHz planar ultrasound transducer, producing multiple focal spots on a target plane. These focal spots generate reverberant shear wave fields detected by the optical coherence tomography (OCT) system. The effectiveness of the multifocal Rev-OCE system in probing mechanical properties with high resolution is demonstrated in layered gelatin phantoms.

Multimodal imaging system combining optical coherence tomography and Brillouin microscopy for neural tube imaging.

To understand the dynamics of tissue stiffness during neural tube formation and closure in a murine model, we have developed a multimodal, coaligned imaging system combining optical coherence tomography (OCT) and Brillouin microscopy. Brillouin microscopy can map the longitudinal modulus of tissue but cannot provide structural images. Thus, it is limited for imaging dynamic processes such as neural tube formation and closure. To overcome this limitation, we have combined Brillouin microscopy and OCT in one coaligned instrument. OCT provided depth-resolved structural imaging with a micrometer-scale spatial resolution to guide stiffness mapping by Brillouin modality. 2D structural and Brillouin frequency shift maps were acquired of mouse embryos at gestational day (GD) 8.5, 9.5, and 10.5 with the multimodal system. The results demonstrate the capability of the system to obtain structural and stiffness information simultaneously.

Longitudinal In Vivo Changes in Radial Peripapillary Capillaries and Optic Nerve Head Structure in Non-Human Primates With Early Experimental Glaucoma.

Purpose: There is conflicting evidence regarding whether a loss of radial peripapillary capillaries (RPCs) precedes neuronal loss in glaucoma. We examined the time course of in vivo changes in RPCs, optic nerve head (ONH) structure, and retinal nerve fiber layer thickness (RNFLT) in experimental glaucoma (EG). Methods: Spectral domain optical coherence tomography images were acquired before and approximately every two weeks after inducing unilateral EG in nine rhesus monkeys to quantify mean anterior lamina cribrosa surface depth (ALCSD), minimum rim width (MRW), and RNFLT. Perfused RPC density was measured from adaptive optics scanning laser ophthalmoscope images acquired on the temporal half of the ONH. The time of first significant change was quantified as when values fell and remained outside of the 95% confidence interval established from control eyes. Results: Mean ALCSD and/or MRW were the first parameters to change in eight EG eyes. RPC density changed first in the ninth. At their first points of change, mean ALCSD posteriorly deformed by 100.2 ± 101.2 µm, MRW thinned by 82.3 ± 65.9 µm, RNFLT decreased by 25 ± 14 µm, and RPC density decreased by 4.5 ± 2.1%. RPC density decreased before RNFL thinning in 5 EG eyes. RNFLT decreased before RPC density decreased in two EG eyes, whereas two EG eyes had simultaneous changes. Conclusions: In most EG eyes, RPC density decreased before (or simultaneous with) a change in RNFLT, suggesting that vascular factors may play a role in axonal loss in some eyes in early glaucoma.

Clinical applications of personalising the neural components of visual image quality metrics for individual eyes.

PURPOSE: Eyecare is evolving increasingly personalised corrections and increasingly personalised evaluations of corrections on-eye. This report describes individualising optical and neural components of the VSX (visual Strehl) metric and evaluates personalisation using two clinical applications. (1) Better understanding visual experience: While VSX tracks visual performance in typical eyes, non-individualised metrics underestimated visual performance in highly aberrated eyes - could this be understood by personalising metrics? (2) Metric-optimised objective spherocylindrical refractions in typical and atypical eyes have used neural weighting functions of typical eyes - will personalisation affect the outcome in clinical 0.25D steps? METHODS: Orientation-specific neural contrast sensitivity was measured in four typical myopic and astigmatic eyes and six eyes with keratoconus. Wavefront error was measured in all eyes while uncorrected and when the keratoconic eyes wore wavefront-guided scleral lenses. Total experiment duration was 24-28 h per subject. Two versions of VSX were calculated for each application: one weighted ocular optics with measured neural contrast sensitivity data from that eye, another weighted optics with a representative neural function of typical eyes. Wavefront-guided corrections were evaluated using the two metric values. Spherocylindrical corrections that optimised each metric were identified. RESULTS: Metric values for keratoconic eyes improved by a mean factor of 1.99 (~0.3 log units) when personalised. Applying this factor to a larger sample of eyes from a previous keratoconus study reconciled dissonances between the percentage of eyes reaching normative best-corrected metric levels and the percentages of eyes reaching normative levels of visual acuity and contrast sensitivity. Spherocylindrical corrections that optimised both versions of VSX were clinically equivalent (mean ± SD Euclidean dioptric difference 0.13 ± 0.18 D). CONCLUSIONS: Personalising visual image quality metrics is beneficial when actual metric values are used (evaluating ophthalmic corrections on-eye against norms) and when fine increments in visual quality are imparted (wavefront-guided corrections). However, partially individualised metrics appear adequate when metrics relatively rank spherocylindrical corrections in 0.25 D steps.

Ultra-fast dynamic line-field optical coherence elastography.

In this work, we present an ultra-fast line-field optical coherence elastography system (LF-OCE) with an 11.5 MHz equivalent A-line rate. The system was composed of a line-field spectral domain optical coherence tomography system based on a supercontinuum light source, Michelson-type interferometer, and a high-speed 2D spectrometer. The system performed ultra-fast imaging of elastic waves in tissue-mimicking phantoms of various elasticities. The results corroborated well with mechanical testing. Following validation, LF-OCE measurements were made in in situ and in in vivo rabbit corneas under various conditions. The results show the capability of the system to rapidly image elastic waves in tissues.

Multimodal high-resolution embryonic imaging with light sheet fluorescence microscopy and optical coherence tomography.

A high-resolution imaging system combining optical coherence tomography (OCT) and light sheet fluorescence microscopy (LSFM) was developed. LSFM confined the excitation to only the focal plane, removing the out of plane fluorescence. This enabled imaging a murine embryo with higher speed and specificity than traditional fluorescence microscopy. OCT gives information about the structure of the embryo from the same plane illuminated by LSFM. The co-planar OCT and LSFM instrument was capable of performing co-registered functional and structural imaging of mouse embryos simultaneously.

Orientation-specific long-term neural adaptation of the visual system in keratoconus.

Eyes with the corneal ectasia keratoconus have performed better than expected (e.g. visual acuity) given their elevated levels of higher-order aberrations that cause rotationally asymmetric retinal blur. Adapted neural processing has been suggested as an explanation but has not been measured across multiple meridional orientations. Using a custom Maxwellian-view laser interferometer to bypass ocular optics, sinusoidal grating neural contrast sensitivity was measured in six eyes (three subjects) with keratoconus and four typical eyes (two subjects) at six spatial frequencies and eight orientations using a two-interval forced-choice paradigm. Total measurement duration was 24 to 28 hours per subject. Neural contrast sensitivity functions of typical eyes agreed with literature and generally showed the oblique effect on a linear-scale and rotational symmetry on a log-scale (rotational symmetry was quantified as the ratio of the minor and major radii of an ellipse fit to all orientations within each spatial frequency; typical eye mean 0.93, median 0.93; where a circle = 1). Mean sensitivities of eyes with keratoconus were 20% to 60% lower (at lower and higher spatial frequencies respectively) than typical eyes. Orientation-specific neural contrast sensitivity functions in keratoconus showed substantial rotational asymmetry (ellipse radii ratio: mean 0.84; median 0.86) and large meridional reductions. The visual image quality metric VSX was used with a permutation test to combine the asymmetric optical aberrations of the eyes with keratoconus and their measured asymmetric neural functions, which illustrated how the neural sensitivities generally mitigated the detrimental effects of the optics.

Automatic Segmentation of Retinal Capillaries in Adaptive Optics Scanning Laser Ophthalmoscope Perfusion Images Using a Convolutional Neural Network.

Purpose: Adaptive optics scanning laser ophthalmoscope (AOSLO) capillary perfusion images can possess large variations in contrast, intensity, and background signal, thereby limiting the use of global or adaptive thresholding techniques for automatic segmentation. We sought to develop an automated approach to segment perfused capillaries in AOSLO images. Methods: 12,979 image patches were extracted from manually segmented AOSLO montages from 14 eyes and used to train a convolutional neural network (CNN) that classified pixels as capillaries, large vessels, background, or image canvas. 1764 patches were extracted from AOSLO montages of four separate subjects, and were segmented manually by two raters (ground truth) and automatically by the CNN, an Otsu's approach, and a Frangi approach. A modified Dice coefficient was created to account for slight spatial differences between the same manually and CNN-segmented capillaries. Results: CNN capillary segmentation had an accuracy (0.94), a Dice coefficient (0.67), and a modified Dice coefficient (0.90) that were significantly higher than other automated approaches (P < 0.05). There were no significant differences in capillary density and mean segment length between manual ground-truth and CNN segmentations (P > 0.05). Conclusions: Close agreement between the CNN and manual segmentations enables robust and objective quantification of perfused capillary metrics. The developed CNN is time and computationally efficient, and distinguishes capillaries from areas containing diffuse background signal and larger underlying vessels. Translational Relevance: This automatic segmentation algorithm greatly increases the efficiency of quantifying AOSLO capillary perfusion images.

Characterization of near-infrared low energy ultra-short laser pulses for portable applications of laser induced breakdown spectroscopy.

We report on the delivery of low energy ultra-short (<1 ps) laser pulses for laser induced breakdown spectroscopy (LIBS). Ultra-short pulses have the advantage of high peak irradiance even at very low pulse energies. This opens the possibility to use compact, rare-earth doped fiber lasers in a portable platform for point detection applications using LIBS for elemental analysis. The use of low energy ultra-short pulses minimizes the generation of a broad continuum background in the emission spectrum, which permits the use of non-gated detection schemes using very simple and compact spectrometers. The pulse energies used to produce high-quality LIBS spectra in this investigation are some of the lowest reported and we investigate the threshold pulse requirements for a number of near IR pulse wavelengths (785-1500 nm) and observe that the pulse wavelength has no effects on the threshold for observation of plasma emission or the quality of the emission spectra obtained.

Ultrafast electronic relaxation and charge-carrier localization in CdS/CdSe/CdS quantum-dot quantum-well heterostructures.

The relaxation and localization times of excited electrons in CdS/CdSe/CdS colloidal quantum wells were measured using subpicosecond spectroscopy. HRTEM analysis and steady-state PL demonstrate a narrow size distribution of 5-6 nm epitaxial crystallites. By monitoring the rise time of the stimulated emission as a function of pump intensity, the relaxation times of the electron from the CdS core into the CdSe well are determined and assigned. Two-component rise times in the stimulated emission are attributed to intraband relaxation of carriers generated directly within the CdSe well (fast component) and charge transfer of core-localized carriers across the CdS/CdSe interface (slow component). This is the first reported observation of simultaneous photon absorption in the core and well of a quantum-dot heterostructure. With increasing pump intensity, the charge-transfer channel between the CdS core CdSe well contributes less to the stimulated emission signal because of filling and saturation of the CdSe well state, making the interfacial charge-transfer component less efficient. The interfacial charge-transfer time of the excited electron was determined from the slow component of the stimulated emission build-up time and is found to have a value of 1.2 ps.

Growth and fragmentation of silver nanoparticles in their synthesis with a fs laser and CW light by photo-sensitization with benzophenone.

The photo-sensitization synthetic technique of making silver nanoparticles using benzophenone is studied using both a laser and a mercury lamp as light sources. The power and irradiation time dependence of the synthesized nanoparticle absorption spectra and their size distribution [as determined by transmission electron microscopy (TEM)] are studied in each method and compared. In the laser synthesis, as either the laser power or the irradiation time increases, the intensity of the surface plasmon resonance absorption at 400 nm is found to increase linearly first, followed by a reduction of the red edge of the plasmon resonance absorption band. The TEM results showed that in the laser synthesis low powers and short irradiation times produce nanoparticles around 20 nm in diameter. Increasing the power or irradiation time produces a second population of nanoparticles with average size of 5 nm in diameter. These small particles are believed to be formed from the surface ablation of the large particles. The surface plasmon absorption band is found to be narrower when the nanoparticles are produced with laser irradiation. Throughout the exposure time with the CW lamp, the plasmon resonance absorption band of the particles formed first grows in intensity, then blue shifts and narrows, and finally red shifts while decreasing in intensity. The TEM results for lamp samples showed particle formation and growth, followed by small nanoparticle formation. The above results are discussed in terms of a mechanism in which, the excited benzophenone forms the ketal radical, which reduces Ag+ in solution and on the Ag nanoparticle surface. As the time of irradiation or the light energy increases the benzophenone is consumed, which is found to be the limiting reagent. This stops the formation of the normal large nanoparticles while their photo-ablation continues to make the small particles.