Absolute retinal blood flowmeter using a laser Doppler velocimeter combined with adaptive optics.

SIGNIFICANCE: The development of a technique allowing for non-invasive measurement of retinal blood flow (RBF) in humans is needed to understand many retinal vascular diseases (pathophysiology) and evaluate treatment with potential improvement of blood flow. AIM: We developed and validated an absolute laser Doppler velocimeter (LDV) based on an adaptive optical fundus camera that provides simultaneously high-definition images of the fundus vessels and absolute maximal red blood cells (RBCs) velocity to calculate the absolute RBF. APPROACH: This new absolute LDV is combined with the adaptive optics (AO) fundus camera (rtx1, Imagine Eyes©, Orsay, France) outside its optical wavefront correction path. A 4-s recording includes 40 images, each synchronized with two Doppler shift power spectra. Image analysis provides a vessel diameter close to the probing beam, and the velocity of the RBCs in the vessels are extracted from the Doppler spectral analysis. A combination of these values gives an average of the absolute RBF. RESULTS: An in vitro experiment consisting of latex microspheres flowing in water through a glass capillary to simulate a blood vessel and in vivo measurements on six healthy humans was done to assess the device. In the in vitro experiment, the calculated flow varied between 1.75 and 25.9 µL / min and was highly correlated (r2 = 0.995) with the flow imposed by a syringe pump. In the in vivo experiment, the error between the flow in the parent vessel and the sum of the flow in the daughter vessels was between -11 % and 36% (mean ± sd, 5.7 ± 18.5 % ). RBF in the main temporal retinal veins of healthy subjects varied between 0.9 and 13.2 µL / min. CONCLUSIONS: The AO LDV prototype allows for the real-time measurement of absolute RBF derived from the retinal vessel diameter and the maximum RBCs velocity in that vessel.

Compact Laser Doppler Flowmeter (LDF) Fundus Camera for the Assessment of Retinal Blood Perfusion in Small Animals.

PURPOSE: Noninvasive techniques for ocular blood perfusion assessment are of crucial importance for exploring microvascular alterations related to systemic and ocular diseases. However, few techniques adapted to rodents are available and most are invasive or not specifically focused on the optic nerve head (ONH), choroid or retinal circulation. Here we present the results obtained with a new rodent-adapted compact fundus camera based on laser Doppler flowmetry (LDF). METHODS: A confocal miniature flowmeter was fixed to a specially designed 3D rotating mechanical arm and adjusted on a rodent stereotaxic table in order to accurately point the laser beam at the retinal region of interest. The linearity of the LDF measurements was assessed using a rotating Teflon wheel and a flow of microspheres in a glass capillary. In vivo reproducibility was assessed in Wistar rats with repeated measurements (inter-session and inter-day) of retinal arteries and ONH blood velocity in six and ten rats, respectively. These parameters were also recorded during an acute intraocular pressure increase to 150 mmHg and after heart arrest (n = 5 rats). RESULTS: The perfusion measurements showed perfect linearity between LDF velocity and Teflon wheel or microsphere speed. Intraclass correlation coefficients for retinal arteries and ONH velocity (0.82 and 0.86, respectively) indicated strong inter-session repeatability and stability. Inter-day reproducibility was good (0.79 and 0.7, respectively). Upon ocular blood flow cessation, the retinal artery velocity signal substantially decreased, whereas the ONH signal did not significantly vary, suggesting that it could mostly be attributed to tissue light scattering. CONCLUSION: We have demonstrated that, while not adapted for ONH blood perfusion assessment, this device allows pertinent, stable and repeatable measurements of retinal blood perfusion in rats.