Glaucoma affects a significant portion of the global population, necessitating regular monitoring of intraocular pressure (IOP). Existing methods are limited by expensive equipment and infrequent measurements. This study proposes an implantable interferometric device for accurate, easy to use, handy and accessible IOP measurement. The device operates through reflection, receiving light from a near-infrared source. A retroreflector directs the light to a spectrometer for analysis. The device consists of a movable membrane and a fixed membrane, with pressure changes leading to spectral variations in the transmitted light. The system achieves a resolution of ~2nm, enabling precise IOP measurement. The design addresses the limitations of current approaches and offers a practical solution for accurate IOP monitoring.
We use optical modelling to reconstruct the pseudophakic eye for 4 different accommodative intra-ocular lens (AIOL) design concepts and map their respective stray light retinal intensity images over a wide range of field of view. The results provide a comparison of stray light performance, both quantitative and qualitative, between the competing AIOL designs. In addition, a deeper analysis of the ray trace offers insights on the various sources of AIOL stray light and leads to operative conclusions on how to minimize AIOL-induced stray light artifacts.
We combined NIR-II illumination at ~1.7 μm with reflectance confocal microscopy and achieved an imaging depth of ~1.3 mm with high spatial resolution in adult mouse brain in vivo, which is 3-4 times deeper than that of conventional confocal microscopy using visible wavelength. We showed that the method can be added as an additional channel to any laser-scanning microscope with low-cost sources and detectors, such as continuous-wave (CW) diode lasers and InGaAs photodiodes. The technique is label-free, simple and requires low illumination power, potentially creating new opportunities for deep tissue imaging in various biological and clinical applications.
We demonstrate three-photon microscopy (3PM) of mouse cerebellum at 1 mm depth by imaging both blood vessels and neurons. We compared 3PM and 2PM in the mouse cerebellum for imaging green (using excitation sources at 1300 nm and 920 nm, respectively) and red fluorescence (using excitation sources at 1680 nm and 1064 nm, respectively). 3PM enabled deeper imaging than 2PM because the use of longer excitation wavelength reduces the scattering in biological tissue and the higher order nonlinear excitation provides better 3D localization. To illustrate these two advantages quantitatively, we measured the signal decay as well as the signal-to-background ratio (SBR) as a function of depth. We performed 2-photon imaging from the brain surface all the way down to the area where the SBR reaches ~ 1, while at the same depth, 3PM still has SBR above 30. The segmented decay curve shows that the mouse cerebellum has different effective attenuation lengths at different depths, indicating heterogeneous tissue property for this brain region. We compared the third harmonic generation (THG) signal, which is used to visualize myelinated fibers, with the decay curve. We found that the regions with shorter effective attenuation lengths correspond to the regions with more fibers. Our results indicate that the widespread, non-uniformly distributed myelinated fibers adds heterogeneity to mouse cerebellum, which poses additional challenges in deep imaging of this brain region.
The attenuation of excitation power reaching the focus is the main issue that limits the depth penetration of highresolution imaging of biological tissue. The attenuation is caused by a combination of tissue scattering and absorption. Theoretical model of the effective attenuation length for in vivo mouse brain imaging has been built based on the data of the absorption of water and blood and the Mie scattering of a tissue-like phantom. Such a theoretical model has been corroborated at a number of excitation wavelengths, such as 800 nm, 1300 nm , and 1700 nm ; however, the attenuation caused by absorption is negligible when compared to tissue scattering at all these wavelength windows. Here we performed in vivo three-photon imaging of Texas Red-stained vasculature in the same mouse brain with different excitation wavelengths, 1700 nm, 1550 nm, 1500 nm and 1450 nm. In particular, our studies include the wavelength regime where strong water absorption is present (i.e., 1450 nm), and the attenuation by water absorption is predicted to be the dominant contribution in the excitation attenuation. Based on the experimental results, we found that the effective attenuation length at 1450 nm is significantly shorter than those at 1700 nm and 1300 nm. Our results confirm that the theoretical model based on tissue scattering and water absorption is accurate in predicting the effective attenuation lengths for in vivo imaging. The optimum excitation wavelength windows for in vivo mouse brain imaging are at 1300 nm and 1700 nm.
Multiphoton fluorescence microscopy is a well-established technique for deep-tissue imaging with subcellular resolution. Three-photon microscopy (3PM) when combined with long wavelength excitation was shown to allow deeper imaging than two-photon microscopy (2PM) in biological tissues, such as mouse brain, because out-of-focus background light can be further reduced due to the higher order nonlinear excitation. As was demonstrated in 2PM systems, imaging depth and resolution can be improved by aberration correction using adaptive optics (AO) techniques which are based on shaping the scanning beam using a spatial light modulator (SLM). In this way, it is possible to compensate for tissue low order aberration and to some extent, to compensate for tissue scattering. Here, we present a 3PM AO microscopy system for brain imaging. Soliton self-frequency shift is used to create a femtosecond source at 1675 nm and a microelectromechanical (MEMS) SLM serves as the wavefront shaping device. We perturb the 1020 segment SLM using a modified nonlinear version of three-point phase shifting interferometry. The nonlinearity of the fluorescence signal used for feedback ensures that the signal is increasing when the spot size decreases, allowing compensation of phase errors in an iterative optimization process without direct phase measurement. We compare the performance for different orders of nonlinear feedback, showing an exponential growth in signal improvement as the nonlinear order increases. We demonstrate the impact of the method by applying the 3PM AO system for in-vivo mouse brain imaging, showing improvement in signal at 1-mm depth inside the brain.
We demonstrate a robust, all-fiber, two-wavelength time-lens source for background-free coherent anti-Stokes Raman scattering (CARS) imaging. The time-lens source generates two picosecond pulse trains simultaneously: one at 1064 nm and the other tunable between 1040 nm and 1075 nm (~ 400 mW for each wavelength). When synchronized to a modelocked Ti:Sa laser, the two wavelengths are used to obtain on- and off-resonance CARS images. Real-time subtraction of the nonresonant background in the CARS image is achieved by the synchronization of the pixel clock and the time-lens source. Background-free CARS imaging of sebaceous glands in ex vivo mouse tissue is demonstrated.
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