(In Reverse Order of publication, most recent first)
Title: Speckle reduction in double-pass retinal images
Authors:Donatus Halpaap, Carlos E. García-Guerra, Meritxell Vilaseca and Cristina Masoller
Abstract: The double pass (DP) technique quantifies the optical quality of the eye by measuring its point spread function. The low reflectivity of the retina requires the use of a high-brightness, point-like illumination source, and thus, DP systems use laser diodes (LDs). However, LDs light produces speckle, and a low-cost solution to reduce speckle is to include a vibrating mirror in the beam path. With the goal of finding an all-optical solution, here we perform a comparative study of the amount of speckle produced by three semiconductor light sources: an LD, a light emitting diode (LED), and a superluminescent diode (SLED). We also compare the results with the speckle reduction that is obtained with a vibrating mirror. We find that the SLED is a good alternative to LD illumination, as the amount of speckle in the image is almost as low as that obtained with an LD and a vibrating mirror in the beam path.
Title: Unsupervised feature extraction of anterior chamber OCT images for ordering and classification
Authors: Pablo Amil, Laura González, Elena Arrondo, Cecilia Salinas, Josep Lluís Guell, Cristina Masoller and Ulrich Parlitz
Abstract: We propose an image processing method for ordering anterior chamber optical coherence tomography (OCT) images in a fully unsupervised manner. The method consists of three steps: Firstly we preprocess the images (filtering the noise, aligning and normalizing the resolution); secondly, a distance measure between images is computed for every pair of images; thirdly we apply a machine learning algorithm that exploits the distance measure to order the images in a two-dimensional plane. The method is applied to a large (~1000) database of anterior chamber OCT images of healthy subjects and patients with angle-closure and the resulting unsupervised ordering and classification is validated by two ophthalmologists.
Title: Energy-Reduced Arrhythmia Termination Using Global Photostimulation in Optogenetic Murine Hearts
Authors: Raúl A. Quiñonez Uribe, Stefan Luther, Laura Diaz-Maue and Claudia Richter
Abstract: Complex spatiotemporal non-linearity as observed during cardiac arrhythmia strongly correlates with vortex-like excitation wavelengths and tissue characteristics. Therefore, the control of arrhythmic patterns requires fundamental understanding of dependencies between onset and perpetuation of arrhythmia and substrate instabilities. Available treatments, such as drug application or high-energy electrical shocks, are discussed for potential side effects resulting in prognosis worsening due to the lack of specificity and spatiotemporal precision. In contrast, cardiac optogenetics relies on light sensitive ion channels stimulated to trigger excitation of cardiomyocytes solely making use of the inner cell mechanisms. This enables low-energy, non-damaging optical control of cardiac excitation with high resolution. Recently, the capability of optogenetic cardioversion was shown in Channelrhodopsin-2 (ChR2) transgenic mice. But these studies used mainly structured and local illumination for cardiac stimulation. In addition, since optogenetic and electrical stimulus work on different principles to control the electrical activity of cardiac tissue, a better understanding of the phenomena behind optogenetic cardioversion is still needed. The present study aims to investigate global illumination with regard to parameter characterization and its potential for cardioversion. Our results show that by tuning the light intensity without exceeding 1.10 mW mm-2, a single pulse in the range of 10–1,000 ms is sufficient to reliably reset the heart into sinus rhythm. The combination of our panoramic low-intensity photostimulation with optical mapping techniques visualized wave collision resulting in annihilation as well as propagation perturbations as mechanisms leading to optogenetic cardioversion, which seem to base on other processes than electrical defibrillation. This study contributes to the understanding of the roles played by epicardial illumination, pulse duration and light intensity in optogenetic cardioversion, which are the main variables influencing cardiac optogenetic control, highlighting the advantages and insights of global stimulation. Therefore, the presented results can be modules in the design of novel illumination technologies with specific energy requirements on the way toward tissue-protective defibrillation techniques.
Title: Three-photon light-sheet fluorescence microscopy
Authors: A. Escobet-Montalbán, F. M. Gasparoli, J. Nylk, P. Liu, Z. Yang, and K. Dholakia
Abstract: We present the first demonstration of three-photon excitation light-sheet fluorescence microscopy. Light-sheet fluorescence microscopy in single- and two-photon modes has emerged as a powerful wide-field, low-photodamage technique for fast volumetric imaging of biological samples. We extend this imaging modality to the three-photon regime, enhancing its penetration depth. Our present study uses a conventional femtosecond pulsed laser at 1000 nm wavelength for the imaging of 450 μm diameter cellular spheroids. In addition, we show, experimentally and through numerical simulations, the potential advantages in three-photon light-sheet microscopy of using propagation-invariant Bessel beams in preference to Gaussian beams.
Title: Wide-field multiphoton imaging through scattering media without correction.
Authors: A. Escobet-Montalbán, R. Spesyvtsev, M. Chen, W. Afshar Saber, M. Andrews, C. Simon Herrington, M. Mazilu and K. Dholakia
Abstract: Optical approaches to fluorescent, spectroscopic, and morphological imaging have made exceptional advances in the last decade. Super-resolution imaging and wide-field multiphoton imaging are now underpinning major advances across the biomedical sciences. While the advances have been startling, the key unmet challenge to date in all forms of optical imaging is to penetrate deeper. A number of schemes implement aberration correction or the use of complex photonics to address this need. In contrast, we approach this challenge by implementing a scheme that requires no a priori information about the medium nor its properties. Exploiting temporal focusing and single-pixel detection in our innovative scheme, we obtain wide-field two-photon images through various turbid media including a scattering phantom and tissue reaching a depth of up to seven scattering mean free path lengths. Our results show that it competes favorably with standard point-scanning two-photon imaging, with up to a fivefold improvement in signal-to-background ratio while showing significantly lower photobleaching.