Bimorph deformable mirror based adaptive optics scanning laser ophthalmoscope for retina imaging in vivo

A bimorph deformable mirror(DM) with a large stroke of more than 30 μm using 35 actuators is presented and characterized for an adaptive optics(AO) confocal scanning laser ophthalmoscope application. Facilitated with a Shack–Hartmann wavefront sensor, the bimorph DM-based AO operates closed-loop AO corrections for human eyes and reduces wavefront aberrations in most eyes to below 0.1 μm rms. Results from living eyes, including one exhibiting ～5D of myopia and ～2D of astigmatism along with notable high-order aberrations, reveal a practical efficient aberration correction and demonstrate a great benefit for retina imaging, including improving resolution, increasing brightness, and enhancing the contrast of images.     

Quick focusing of imaging optics using micromachined adaptive mirrors

Inexpensive micromachined varifocal mirrors, providing a very wide range of focal distance control are described. The focal distance of a mirror with a clear aperture of 1 cm can be controlled in the range ∞ to +0.25 m (four diopters) in a frequency band up to 75 Hz by applying control voltages in the range of 0 to 200 V. Applications for quick focus control in a CCD camera and for direct correction of the accommodation depth of a human eye are reported.     

Retinal imaging with adaptive optics

Imaging the human retina in vivo is affected by the eye's natural aberrations, which limit the resolution of retinal images. Measuring these aberrations, including the high order ones, is possible using wavefront sensing techniques. A review of the rapid progress in this field is given. Once the aberrations are known, adaptive optics methods, developed for astronomical observing in the past 15 years, can be applied in order either to improve retinal imaging or to give hyper-vision to the subject. Progress in this domain is reviewed, and some original results are reported with a new instrument. Future applications are discussed, including a possible three-dimensional, high-resolution method to image the human retina in vivo.     

Stokes vector analysis of adaptive optics images of the retina

A high-resolution Stokes vector imaging polarimeter was developed to measure the polarization properties at the cellular level in living human eyes. The application of this cellular level polarimetric technique to in vivo retinal imaging has allowed us to measure depolarization in the retina and to improve the retinal image contrast of retinal structures based on their polarization properties.     

In vivo imaging of mice auricle vessels using adaptive optical confocal fluorescence microscope

Facilitated with stochastic parallel gradient descent(SPGD) algorithm for wavefront sensorless correcting aberrations, an adaptive optics(AO) confocal fluorescence microscopy is developed and used to record fluorescent signals in vivo. Vessels of mice auricle at 80, 100 and 120 μm depth are obtained, and image contrast and fluorescence intensity are significantly improved with AO correction. The typical 10%–90% rise-time of the metric value measured is 5.0 s for a measured close-loop bandwidth of 0.2 Hz. Therefore, the AO confocal microscopy implemented with SPGD algorithm for robust AO corrections will be a powerful tool for study of vascular dynamics in future.     

In vivo imaging of blood flow in the mouse Achilles tendon using high-frequency ultrasound

Objective

Achilles tendinitis is a common clinical problem with many treatment modalities, including physical therapy, exercise and therapeutic ultrasound. However, evaluating the effects of current therapeutic modalities and studying the therapeutic mechanism(s) in vivo remains problematic. In this study, we attempted to observe the morphology and microcirculation changes in mouse Achilles tendons between pre- and post-treatment using high-frequency (25 MHz) ultrasound imaging. A secondary aim was to assess the potential of high-frequency ultrasound in exploring therapeutic mechanisms in small-animal models in vivo.

Methods

A collagenase-induced mouse model of Achilles tendinitis was adopted, and 5 min treatment of continuous-mode low-frequency (45 kHz) ultrasound with 47 mW/cm² maximum intensity and 16.3 cm² effective beam radiating area was applied. The B-mode images showed no focal hypoechoic regions in normal Achilles tendons either pre- or post-treatment. The Doppler power energy and blood flow rate were measured within the peritendinous space of the Achilles tendon.

Conclusion

An increase in the microcirculation was observed soon after the low-frequency ultrasound treatment, which was due to immediate induction of vascular dilatation. The results suggest that applying high-frequency Doppler imaging to small-animal models will be an invaluable aid in explorations of the therapeutic mechanism(s). Our future work includes using imaging to assess microcirculation changes in tendinitis between before and after treatment over a long time period, which is expected to yield useful physiological data for future human studies.     

Simulated human eye retina adaptive optics imaging system based on a liquid crystal on silicon device

In order to obtain a clear image of the retina of model eye, an adaptive optics system used to correct the wave-front error is introduced in this paper. The spatial light modulator that we use here is a liquid crystal on a silicon device instead of a conversional deformable mirror. A paper with carbon granule is used to simulate the retina of human eye. The pupil size of the model eye is adjustable (3--7mm). A Shack-Hartman wave-front sensor is used to detect the wave-front aberration. With this construction, a value of peak-to-valley is achieved to be 0.086Λ, where Λ is wavelength. The modulation transfer functions before and after corrections are compared. And the resolution of this system after correction (69lp/m) is very close to the diffraction limit resolution. The carbon granule on the white paper which has a size of 4.7μm is seen clearly. The size of the retina cell is between 4 and 10μm. So this system has an ability to image the human eye's retina.     

Adaptive optics optical coherence tomography for retina imaging

When optical coherence tomography (OCT) is used for human retina imaging, its transverse resolution is limited by the aberrations of human eyes. To overcome this disadvantage, a high resolution imaging system for living human retina, which consists of a time domain OCT system and a 37-elements adaptive optics (AO) system, has been developed. The AO closed loop rate is 20 frames per second, and the OCT has a 6.7-μm axial resolution. In this paper, this system is introduced and the high resolution imaging results for retina are presented.     

基于成像自适应光学的水下成像系统研究

水中湍流是影响水下光学系统性能的重要的潜在因素。用波前检测的方法对水中湍流对激光束传输的影响进行实验研究,测量了实验条件下水中湍流产生的波前畸变的大小,对波前畸变进行了Zernike分析,还对波前补偿自适应系统在水下成像的应用进行了探索。结果表明:水中湍流引起的激光束波前的起伏降低了图像的分辨率,自适应光学系统在其校正范围内能有效地校正畸变,提高图像质量。     

In vivo photoacoustic chorioretinal vascular imaging in albino mouse

Photoacoustic ophthalmoscopy(PAOM) is a novel imaging modality, which is capable of non-invasively detecting optical absorption properties in the retina. We visualize the microvasculature in retina and choroid in albino mouse using PAOM guided by spectral-domain optical coherence tomography. Since albino mouse characterizes by lacking melanin in retinal pigment epithelium(RPE), PAOM illumination laser can penetrate through the RPE onto choroid, and consequently provides volumetric visualization of chorioretinal vasculatures as a result of strong hemoglobin optical absorption. The high-quality chorioretinal microvascular imaging acquired by PAOM implies its great potential in understanding pathological mechanisms and developing therapeutic strategies for major chorioretinal diseases that correlate with vascular disorders.     

High resolution mosaic image of capillaries in human retina by adaptive optics

Adaptive optics (AO) has been proved as a powerful means for high resolution imaging of human retina. Because of the pixel number of charge-coupled device (CCD) camera, the field of view is limited to 1°. In order to have image of capillaries around vivo human fovea, we use mosaic method to obtain high resolution image in area of 6°×6°. Detailed structures of capillaries around fovea with resolution of 2.3 μm are clearly shown. Comparison shows that this method has a much higher resolution than current clinic retina imaging methods.     

High resolution mosaic image of capillaries in human retina by adaptive optics

Adaptive optics (AO) has been proved as a powerful means for high resolution imaging of human retina.Because of the pixel number of charge-coupled device (CCD) camera, the field of view is limited to 1°.In order to have image of capillaries around vivo human fovea, we use mosaic method to obtain high resolution image in area of 6°× 6°. Detailed structures of capillaries around fovea with resolution of 2.3μm are clearly shown. Comparison shows that this method has a much higher resolution than current clinic retina imaging methods.     

In situ roughness monitoring of sputtered Pt thin film under dynamic turbulence using adaptive optics

A new optical monitoring system for rapid and in situ surface roughness measurement of Pt film on silicon is developed in this study. The in-process measurement is achieved by combining an optical probe of laser-scattering phenomena and adaptive optics for aberration correction. Platinum (Pt) thin film is selected due to the extensive utilization in semiconductor industry and excellent chemical inertness. The aim of this study was to demonstrate the necessity for adaptive optics (AO) compensation in regions containing room-temperature turbulences. Measurement results of eight Pt films (roughness ranging from 58 to 83 nm) sputtered on top of P-type silicon wafer demonstrate excellent correlation between the peak power and average roughness with a correlation coefficient (R²) of 0.9962 and a trend equation for predicting the surface roughness of Pt thin films is obtained as y = 9E07x^−3.783. Roughness average (Ra) of Pt thin films (x) of can be directly determined from the peak power (y) using the proposed method under dynamic disturbance. Furthermore, the proposed AO-assisted system is in good agreement with stylus method and less than 1.1% error values are obtained for the aforementioned average sample roughness.     

液晶自适应光学视网膜校正成像技术研究

为了实现对人眼视网膜的高分辨率成像,解决偏振能量损失、成像视场小和普适性差等问题,对液晶自适应光学技术及其在人眼视网膜成像中的应用进行了研究。通过开环光路的设计方案,避免了闭环液晶自适应系统的偏振光能量损失;在光路中加入可变视场光阑,利用小视场照明进行波前探测、大视场照明进行像差校正和成像的方法扩大了成像视场;使用脉冲光照明的方案减小曝光量;通过偏振光照明提高能量利用率、等效无穷远视标配合补偿镜以及改进后的视标提高盯视稳定性等一系列方法,提高系统普适性。校正后成像的清晰度和对比度获得了明显提高;高分辨率眼底成像视场直径从200 μm扩大到500 μm;曝光量减小到原来的1/2~1/3;对前期难以获得清晰成像的样本,取得了效果良好的视网膜视觉细胞自适应图像。     

In vivo imaging of intrinsic optical signals in chicken retina with functional optical coherence tomography

Visually evoked intrinsic optical signals (IOSs) were measured in vivo for the first time to our knowledge from all retina layers of the chicken retina with a combined functional optical coherence tomography and electroretinography (ERG) system. IOS traces were recorded from a small volume in the retina with 3.5 μm axial resolution and 7 ms time resolution. Comparison of the IOS and ERG traces shows a correlation between the positive and negative IOS measured from different retinal layers and the timing of the a and b waves in the ERG recording.     

Rapid in-process measurement of surface roughness using adaptive optics

We present an in-process measurement of surface roughness by combining an optical probe of laser-scattering phenomena and adaptive optics for aberration correction. Measurement results of five steel samples with a roughness ranging from 0.2 to 3.125 μm demonstrate excellent correlation between the peak power and average roughness with a correlation coefficient (R(2)) of 0.9967. The proposed adaptive-optics-assisted system is in good agreement with the stylus method, and error values of less than 8.7% are obtained for average sample roughness in the range of 0.265 to 1.119 μm. The proposed system can be used as a rapid in-process roughness monitor/estimator to further increase the precision and stability of manufacturing processes in situ.     

Solar adaptive optics system using an electromagnetic deformable mirror

A solar adaptive optics system has been improved by using a high-speed electromagnetic deformable mirror and adopting a modified sum-of-absolute-differences algorithm in wavefront sensing. Results of laboratory experiments clearly show that the use of the mirror raises the temporal performance of the system. In solar observations, wavefront compensation using solar granules as a target is realized.     

In vivo hypoxia characterization using blood oxygen level dependent magnetic resonance imaging in a preclinical glioblastoma mouse model

PurposeHypoxia measurements can provide crucial information regarding tumor aggressiveness, however current preclinical approaches are limited. Blood oxygen level dependent (BOLD) Magnetic Resonance Imaging (MRI) has the potential to continuously monitor tumor pathophysiology (including hypoxia). The aim of this preliminary work was to develop and evaluate BOLD MRI followed by post-image analysis to identify regions of hypoxia in a murine glioblastoma (GBM) model.MethodsA murine orthotopic GBM model (GL261-luc2) was used and independent images were generated from multiple slices in four different mice. Image slices were randomized and split into training and validation cohorts. A 7 T MRI was used to acquire anatomical images using a fast-spin-echo (FSE) T2-weighted sequence. BOLD images were taken with a T2*-weighted gradient echo (GRE) and an oxygen challenge. Thirteen images were evaluated in a training cohort to develop the MRI sequence and optimize post-image analysis. An in-house MATLAB code was used to evaluate MR images and generate hypoxia maps for a range of thresholding and ΔT2* values, which were compared against respective pimonidazole sections to optimize image processing parameters. The remaining (n = 6) images were used as a validation group. Following imaging, mice were injected with pimonidazole and collected for immunohistochemistry (IHC). A test of correlation (Pearson's coefficient) and agreement (Bland-Altman plot) were conducted to evaluate the respective MRI slices and pimonidazole IHC sections.ResultsFor the training cohort, the optimized parameters of “thresholding” (20 ≤ T2* ≤ 35 ms) and ΔT2* (±4 ms) yielded a Pearson's correlation of 0.697. These parameters were applied to the validation cohort confirming a strong Pearson's correlation (0.749) when comparing the respective analyzed MR and pimonidazole images.ConclusionOur preliminary study supports the hypothesis that BOLD MRI is correlated with pimonidazole measurements of hypoxia in an orthotopic GBM mouse model. This technique has further potential to monitor hypoxia during tumor development and therapy.     

Low-order adaptive optics: a possible use in underwater imaging?

The possibility of applying adaptive optical technology to underwater imaging is discussed. An introduction to the history and problems associated with imaging through turbulence is outlined. Trends in low-cost adaptive optical technology and results from a system applied to the correction of water generated turbulence are presented. The aim of this paper is to demonstrate the current solutions used in the correction of atmospheric turbulence in the hope of applying these same techniques to underwater imaging and communications.     

Retina imaging system with adaptive optics for the eye with or without myopia

An adaptive optics system for the retina imaging is introduced in the paper. It can be applied to the eye with myopia from 0 to 6 diopters without any adjustment of the system. A high-resolution liquid crystal on silicon (LCOS) device is used as the wave-front corrector. The aberration is detected by a Shack-Harmann wave-front sensor (HASO) that has a Root Mean Square (RMS) measurement accuracy of λ/100 (λ = 0.633 μm). And an equivalent scale model eye is constructed with a short focal length lens (∼18 mm) and a diffuse reflection object (paper screen) as the retina. By changing the distance between the paper screen and the lens, we simulate the eye with larger diopters than 5 and the depth of field. The RMS value both before and after correction is obtained by the wave-front sensor. After correction, the system reaches the diffraction-limited resolution approximately 230 cycles/mm at the object space. It is proved that if the myopia is smaller than 6 diopters and the depth of field is between −40 and +50 mm, the system can correct the aberration very well.