KEYWORDS: Retina, Angiography, Optical coherence tomography, Visualization, Laser coagulation, Pathogens, 3D modeling, Animal model studies, Pathophysiology, Drug development
To understand the pathogenesis of ophthalmic disease, utilizing small animal models such as mouse is necessary because of their ease of maintenance and availability. For identifying pathophysiology and drug development of retinal diseases in mouse model, optical coherence tomography angiography (OCTA) is promising imaging modality visualizing not only microstructure but also microvasculature. In this study, we serially imaged 3D structure and angiography of laser-induced choroidal neovascularization (CNV) in the mouse retina with/without anti-VEGF treatment. Also, the volume changes of CNV and avascular region in choroid layer are measured for identifying effects of anti-VEGF.
A lab-built high-speed OCTA prototype using the wavelength-swept laser centered at 1040 nm with 230 kHz A-scan rate acquired 3-D volumetric data consisted of 1024 x 1024 x 3 A-scans. The OCTA scanned 1.7 mm x 1.7 mm area around ONH. For obtaining angiography, amplitude decorrelation from 3 consecutive B-scans at each position was generated. Seven days after the laser photocoagulation at mouse retina for generation of the laser-induced CNV, intravitreal administration of Fc and VEGF-Trap was given in the therapeutic arm. The OCTA were performed at 6, 14, 21 and 35 days after laser photocoagulation. Vasculatures of inner retina, outer retina and choroid layers were separately visualized after RPE flattening and layer segmentation. To investigate therapeutic effects of anti-VEGF treatment, the relative area and volume of CNV in outer retina layer is measured. Also, total volume of avascular zone surrounding the laser injury site in choroid layer is also analyzed.
OCT angiography (OCTA) has recently garnered immense interest in clinical ophthalmology, permitting ocular vasculature to be viewed in exquisite detail, in vivo, and without the injection of exogenous dyes. However, commercial OCTA systems provide little information about actual erythrocyte speeds; instead, OCTA is typically used to visualize the presence and/or absence of vasculature. This is an important limitation because in many ocular diseases, including diabetic retinopathy (DR) and age-related macular degeneration (AMD), alterations in blood flow, but not necessarily only the presence or absence of vasculature, are thought to be important in understanding pathogenesis. To address this limitation, we have developed an algorithm, variable interscan time analysis (VISTA), which is capable of resolving different erythrocyte speeds. VISTA works by acquiring >2 repeated B-scans, and then computing multiple OCTA signals corresponding to different effective interscan times. The OCTA signals corresponding to different effective interscan times contain independent information about erythrocyte speed. In this study we provide a theoretical overview of VISTA, and investigate the utility of VISTA in studying blood flow alterations in ocular disease. OCTA-VISTA images of eyes with choroidal neovascularization, geographic atrophy, and diabetic retinopathy are presented.
The purpose of this study is to evaluate the suitability of five different anesthetic protocols (isoflurane, isoflurane–xylazine, pentobarbital, ketamine–xylazine, and ketamine–xylazine–vecuronium) for functional blood flow imaging in the rat eye. Total retinal blood flow was measured at a series of time points using an ultrahigh-speed Doppler OCT system. Additionally, each anesthetic protocol was qualitatively evaluated according to the following criteria: (1) time-stability of blood flow, (2) overall rate of blood flow, (3) ocular immobilization, and (4) simplicity. We observed that different anesthetic protocols produced markedly different blood flows. Different anesthetic protocols also varied with respect to the four evaluated criteria. These findings suggest that the choice of anesthetic protocol should be carefully considered when designing and interpreting functional blood flow studies in the rat eye.
Since the first demonstration of swept source optical coherence tomography (SS-OCT) imaging using widely tunable micro-electromechanical systems vertical cavity surface-emitting lasers (MEMS-VCSELs) in 2011, VCSEL-based SSOCT has advanced in both device and system performance. These advances include extension of MEMS-VCSEL center wavelength to both 1060nm and 1300nm, improved tuning range and tuning speed, new SS-OCT imaging modes, and demonstration of the first electrically pumped devices. Optically pumped devices have demonstrated continuous singlemode tuning range of 150nm at 1300nm and 122nm at 1060nm, representing a fractional tuning range of 11.5%, which is nearly a factor of 3 greater than the best reported MEMS-VCSEL tuning ranges prior to 2011. These tuning ranges have also been achieved with wavelength modulation rates of >500kHz, enabling >1 MHz axial scan rates. In addition, recent electrically pumped devices have exhibited 48.5nm continuous tuning range around 1060nm with 890kHz axial scan rate, representing a factor of two increase in tuning over previously reported electrically pumped MEMS-VCSELs in this wavelength range. New imaging modes enabled by optically pumped devices at 1060nm and 1300nm include full eye length imaging, pulsatile Doppler blood flow imaging, high-speed endoscopic imaging, and hand-held wide-field retinal imaging.
In the last 2 years, the field of micro-electro-mechanical systems tunable vertical cavity surface-emitting lasers (MEMS-VCSELs)
has seen dramatic improvements in laser tuning range and tuning speed, along with expansion into unexplored
wavelength bands, enabling new applications. This paper describes the design and performance of high-speed ultra-broad
tuning range 1050nm and 1310nm MEMS-VCSELs for medical imaging and spectroscopy. Key results include
achievement of the first MEMS-VCSELs at 1050nm and 1310nm, with 100nm tuning demonstrated at 1050nm and
150nm tuning at shown at 1310nm. The latter result represents the widest tuning range of any MEMS-VCSEL at any
wavelength. Wide tuning range has been achieved in conjunction with high-speed wavelength scanning at rates beyond 1
MHz. These advances, coupled with recent demonstrations of very long MEMS-VCSEL dynamic coherence length,
have enabled advancements in both swept source optical coherence tomography (SS-OCT) and gas spectroscopy.
VCSEL-based SS-OCT at 1050nm has enabled human eye imaging from the anterior eye through retinal and choroid
layers using a single instrument for the first time. VCSEL-based SS-OCT at 1310nm has enabled real-time 3-D SS-OCT
imaging of large tissue volumes in endoscopic settings. The long coherence length of the VCSEL has also enabled, for
the first time, meter-scale SS-OCT applicable to industrial metrology. With respect to gas spectroscopy, narrow dynamic
line-width has allowed accurate high-speed measurement of multiple water vapor and HF absorption lines in the 1310nm
wavelength range, useful in gas thermometry of dynamic combustion engines.
Measuring retinal hemodynamics in response to flicker stimulus is important for investigating pathophysiology in small animal models of diabetic retinopathy, because a reduction in the hyperemic response is thought to be one of the earliest changes in diabetic retinopathy. In this study, we investigated functional imaging of retinal hemodynamics in response to flicker stimulus in the rat retina using an ultrahigh speed spectral / Fourier domain OCT system at 840nm with an axial scan rate of 244kHz. At 244kHz the nominal axial velocity range that could be measured without phase wrapping was +/-37.7mm/s. Pulsatile total retinal arterial blood flow as a function of time was measured using an en face Doppler approach where a 200μm × 200μm area centered at the central retinal artery was repeatedly raster scanned at a volume acquisition rate of 55Hz. Three-dimensional capillary imaging was performed using speckle decorrelation which has minimal angle dependency compared to other angiography techniques based on OCT phase information. During OCT imaging, a flicker stimulus could be applied to the retina synchronously by inserting a dichroic mirror in the imaging interface. An acute transient increase in total retinal blood flow could be detected. At the capillary level, an increase in the degree of speckle decorrelation in capillary OCT angiography images could also be observed, which indicates an increase in the velocity of blood at the capillary level. This method promises to be useful for the investigation of small animal models of ocular diseases.
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