Optical microangiography of retina and choroid and measurement of total retinal blood flow in mice

Zhongwei Zhi, Xin Yin, Suzan Dziennis, Tomasz Wietecha, Kelly L Hudkins, Charles E Alpers, Ruikang K Wang, Zhongwei Zhi, Xin Yin, Suzan Dziennis, Tomasz Wietecha, Kelly L Hudkins, Charles E Alpers, Ruikang K Wang

Abstract

We present a novel application of optical microangiography (OMAG) imaging technique for visualization of depth-resolved vascular network within retina and choroid as well as measurement of total retinal blood flow in mice. A fast speed spectral domain OCT imaging system at 820nm with a line scan rate of 140 kHz was developed to image the posterior segment of eyes in mice. By applying an OMAG algorithm to extract the moving blood flow signals out of the background tissue, we are able to provide true capillary level imaging of the retinal and choroidal vasculature. The microvascular patterns within different retinal layers are presented. An en face Doppler OCT approach [Srinivasan et al., Opt Express 18, 2477 (2010)] was adopted for retinal blood flow measurement. The flow is calculated by integrating the axial blood flow velocity over the vessel area measured in an en face plane without knowing the blood vessel angle. Total retinal blood flow can be measured from both retinal arteries and veins. The results indicate that OMAG has the potential for qualitative and quantitative evaluation of the microcirculation in posterior eye compartments in mouse models of retinopathy and neovascularization.

Keywords: (170.3880) Medical and biological imaging; (170.4500) Optical coherence tomography.

Figures

Fig. 1
Fig. 1
Schematic of the high speed spectral domain OCT/OMAG system. CMOS: line scan camera, PC: polarization controller, SLD: superluminescent diode.
Fig. 2
Fig. 2
Schematic of en face Doppler approach for flow measurement. (A) In conventional Doppler OCT methods, the blood vessel angle ϴ is required to compute absolute velocity values Vabs. Total blood flow is calculated by multiplying with Vabs the cross-sectional area of the vessel S'. (B) In En faceDoppler method, total blood flow is computed by simply integrating the axial velocity components Vz over the en face cross-section S that intercepts the vessel.
Fig. 3
Fig. 3
OCT imaging of morphology in mouse posterior eye. A) OCT fundus image of mouse posterior eye, arrows point to optic nerve fiber bundles. B) Recognition of all the mouse retinal layers and choroid layer. NFL: nerve fiber layer, GCL: ganglion cell layer, IPL: inner plexiform layer, INL: inner nuclear layer, OPL: outer plexiform layer, ONL: outer nuclear layer, ELM: external limiting membrane, IS/OS: junction between the inner and outer segment of the photoreceptors, RPE: retinal pigment epithelium layer, CH: choroid. C) OCT section across the periphery retina. D) OCT section across the central retina (optic nerve head *) and E) corresponding blood flow image. Scale bar = 200 µm.
Fig. 4
Fig. 4
(A) Color coded depth projection map of the retinal vascular network, blue: superficial, green: deep. (B-D) Vascular perfusion maps within different retinal layers in mouse eye after segmentation, where (B) is microvasculature within NFL and GCL, A: artery, V: vein, (C) within IPL, and (D) within OPL. Image size: 2x2 mm2
Fig. 5
Fig. 5
Vascular perfusion map of choroid in mice eye. Arrows points to long posterior ciliary artery (LPCA) entering the choroid. ONH: optic nerve head.
Fig. 6
Fig. 6
Measurement of total retinal blood flow. (A) Angiography obtained with UHS-OMAG. (B) Maximum projection of bi-directional axial flow velocity obtained with Phase-resolved Doppler OCT analysis for region marked by yellow square in (A). Red: artery, green: vein. (C, D) En faceplane view of the axial blood flow velocity at ~500 µm depth, where white squares denote the integration of arterial flow and yellow squares denote the integration of venous flow. Scale bar = 100 µm.

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