Laser Doppler Holography Research Articles

Multiwavelength laser doppler holography (MLDH) in spatiotemporal optical coherence tomography (STOC-T)

Spatiotemporal optical coherence tomography (STOC-T) is the novel modality for high-speed, crosstalk- and aberration-free volumetric imaging of biological tissue in vivo. STOC-T extends the Fourier-Domain holographic Optical Coherence Tomography by the spatial phase modulation that enables the reduction of spatial coherence of the tunable laser. By reducing the spatial coherence of the laser, we suppress coherent noise, and, consequently, improve the imaging depth. Furthermore, we remove geometrical aberrations computationally in postprocessing. We recently demonstrated high-speed, high-resolution STOC-T of human retinal imaging in vivo. Here, we show that the dataset produced by STOC-T can be processed differently to reveal blood flow in the human retina in vivo. To render the blood flow, we first pre-process STOC-T holographic data to access the approximated information about the Doppler-shifted optical field backscattered from the sample. Then, we analyze it using methods from the laser Doppler flowmetry, namely, by analyzing the Doppler broadening caused by moving light scatterers (red blood cells). However, contrary to conventional approaches, we use multiple illumination wavelengths. This enables us to render the structural volumetric and blood flow images from the same dataset concurrently. Our method, denoted as multiwavelength laser Doppler holography (MLDH), links laser Doppler flowmetry with multiwavelength holographic detection to enable noninvasive visualization and possible blood flow quantification at different human retina layers at high speeds and high transverse resolution in vivo.

Retinal blood flow imaging with combined full-field swept-source optical coherence tomography and laser Doppler holography.

Full-field swept-source optical coherence tomography (FF-SS-OCT) and laser Doppler holography (LDH) are two holographic imaging techniques presenting unique capabilities for ophthalmology. We report on interlaced FF-SS-OCT and LDH imaging with a single instrument. Effectively, retinal blood flow and pulsation could be quasi-simultaneously monitored. This instrument holds potential for a wide scope of ophthalmic applications.

Measurement of the arterial wall elasticity using the velocity of flexural waves in retinal blood vessels

Arterial stiffness (typically a Young modulus of 100 kPa) is known to be an indicator of the cardiovascular health of a patient. It is commonly used in clinics to prevent and anticipate cardiovascular diseases. The existing methods allowing to measure the arterial wall elasticity measuring the velocity of the natural pulse wave lack of accuracy. We propose here a method consisting in using laser Doppler holography experiments to estimate the arterial wall elasticity. Laser Doppler holography experiments are performed on patients and noise correlation algorithms are used on these data to measure the wave velocity. We measured separately the wavelength and the frequency of elastic waves, allowing to calculate their velocity. Due to the temporal resolution of the raw data and to the way the data are processed, the waves we detected are not the classical axisymmetric pulse wave. The physics of guided waves indicates that both antisymmetric (called A0) and symmetric (called S0) waves can propagate in tubes, and in this case only antisymmetric flexural waves are detected. They are both sensitive to the arterial wall elasticity, and at low frequencies, flexural (or antisymmetric) waves are much slower than the axisymmetric pulse wave, about 3 mm/s, allowing to increase greatly the measurement accuracy. When knowing the diameter of the blood vessel, for the first time it is possible to retrieve the arterial wall elasticity thanks to the antisymmetric wave velocity measurement.

Retinal blood flow reversal quantitatively monitored in out-of-plane vessels with laser Doppler holography

Laser Doppler holography is a planar blood flow imaging technique recently introduced in ophthalmology to image human retinal and choroidal blood flow non-invasively. Here we present a digital method based on the Doppler spectrum asymmetry that reveals the local direction of blood flow with respect to the optical axis in out-of-plane vessels. This directional information is overlaid on standard grayscale blood flow images to depict flow moving towards the camera in red and flow moving away from the camera in blue, as in ultrasound color Doppler imaging. We show that thanks to the strong contribution of backscattering to the Doppler spectrum in out-of-plane vessels, the local axial direction of blood flow can be revealed with a high temporal resolution, which enables us to evidence pathological blood flow reversals. We also demonstrate the use of optical Doppler spectrograms to quantitatively monitor retinal blood flow reversals.

Laser Doppler holography of the anterior segment for blood flow imaging, eye tracking, and transparency assessment

Laser Doppler holography (LDH) is a full-field blood flow imaging technique able to reveal human retinal and choroidal blood flow with high temporal resolution. We here report on using LDH in the anterior segment of the eye without making changes to the instrument. Blood flow in the bulbar conjunctiva and episclera as well as in corneal neovascularization can be effectively imaged. We additionally demonstrate simultaneous holographic imaging of the anterior and posterior segments by simply adapting the numerical propagation distance to the plane of interest. We used this feature to track the movements of the retina and pupil with high temporal resolution. Finally, we show that the light backscattered by the retina can be used for retro-illumination of the anterior segment. Hence digital holography can reveal opacities caused by absorption or diffusion in the cornea and eye lens.

Reverse contrast laser Doppler holography for lower frame rate retinal and choroidal blood flow imaging.

Laser Doppler holography (LDH) is an interferometric blood flow imaging technique based on full-field measurements of the Doppler spectrum. LDH has so far been demonstrated in the retina with ultrafast cameras, typically at 75 kHz. We show here that a similar method can be implemented with camera frame rates 10 times slower than before. Due to energy conservation, low and high frequency local power Doppler signals have opposite variations, and a simple contrast inversion of the low frequency power Doppler reveals fast blood flow beyond the camera detection bandwidth for conventional laser Doppler measurements. Relevant blood flow variations and color composite power Doppler images can be obtained with camera frame rates down to a few kHz.

Spatio-temporal filtering in laser Doppler holography for retinal blood flow imaging.

Laser Doppler holography (LDH) is a full-field interferometric imaging technique recently applied in ophthalmology to measure blood flow, a parameter of high clinical interest. From the temporal fluctuations of digital holograms acquired at ultrafast frame rates, LDH reveals retinal and choroidal blood flow with a few milliseconds of temporal resolution. However, LDH experiences difficulties to detect slower blood flow as it requires to work with low Doppler frequency shifts which are corrupted by eye motion. We here demonstrate the use of a spatio-temporal decomposition adapted from Doppler ultrasound that provides a basis appropriate to the discrimination of blood flow from eye motion. A singular value decomposition (SVD) can be used as a simple, robust, and efficient way to separate the Doppler fluctuations of blood flow from those of strong spatial coherence such as eye motion. We show that the SVD outperforms the conventional Fourier based filter to reveal slower blood flow, and dramatically improves the ability of LDH to reveal vessels of smaller size or with a pathologically reduced blood flow.

Waveform analysis of human retinal and choroidal blood flow with laser Doppler holography.

Laser Doppler holography was introduced as a full-field imaging technique to measure blood flow in the retina and choroid with an as yet unrivaled temporal resolution. We here investigate separating the different contributions to the power Doppler signal in order to isolate the flow waveforms of vessels in the posterior pole of the human eye. Distinct flow behaviors are found in retinal arteries and veins with seemingly interrelated waveforms. We demonstrate a full field mapping of the local resistivity index, and the possibility to perform unambiguous identification of retinal arteries and veins on the basis of their systolodiastolic variations. Finally we investigate the arterial flow waveforms in the retina and choroid and find synchronous and similar waveforms, although with a lower pulsatility in choroidal arteries. This work demonstrates the potential held by laser Doppler holography to study ocular hemodynamics in healthy and diseased eyes.

Choroidal vasculature imaging with laser Doppler holography.

The choroid is a highly vascularized tissue supplying the retinal pigment epithelium and photoreceptors. Its implication in retinal diseases is gaining increasing interest. However, investigating the anatomy and flow of the choroid remains challenging. Here we show that laser Doppler holography provides high-contrast imaging of choroidal vessels in humans, with a spatial resolution comparable to state-of-the-art indocyanine green angiography and optical coherence tomography. Additionally, laser Doppler holography contributes to sort out choroidal arteries and veins by using a power Doppler spectral analysis. We thus demonstrate the potential of laser Doppler holography to improve our understanding of the anatomy and flow of the choroidal vascular network.

Laser Doppler holography reveals retinal blood flow in humans

Laser Doppler holography reveals retinal blood flow in humans

Blood flow imaging in zebrafish by laser doppler digital holography.

Microvessel blood flow imaging techniques are widely used in biomedical research and clinical diagnostics where many diseases have a vascular etiology or involvement. For testing purposes, zebrafish embryo provides an ideal animal model to achieve high-resolution imaging of superficial and deeply localized vessels. Moreover, the study of the formation of a closed circulatory system in vertebrates is a topic of recent interest in biophysics. However, most of the existing techniques are invasive due to the use of a contrast agent for imaging purposes. Recent developments in Digital Holography and Laser Doppler Holography techniques can be considered to alleviate this issue. Laser Doppler holography and transmission microscopy can be coupled to analyze blood flow in fish embryos by adapting a laser Doppler holographic setup to a standard bio-microscope: the two beams of the holographic interferometer (illumination of the object and reference), whose frequency offset is controlled, were addressed to the microscope by optical fibers. Multimodal acquisition and analysis of the data is made by acting on the frequency offset of the two beams, and on the location of the Fourier space filtered zone. In this work, we show that it is possible to select the signal of moving scatterers, and to image Red Blood Cells (RBCs) and blood vessels. Individual RBCs are imaged, and movies showing the RBC motion are obtained. Microsc. Res. Tech. 81:153-161, 2018. © 2016 Wiley Periodicals, Inc.

Short-time Fourier transform laser Doppler holography

We report a demonstration of laser Doppler holography at a sustained acquisition rate of 250 Hz on a 1 Megapixel complementary metal-oxide-semiconductor (CMOS) sensor array and image display at 10 Hz frame rate. The holograms are optically acquired in off-axis configuration, with a frequency-shifted reference beam. Wide-field imaging of optical fluctuations in a 250 Hz frequency band is achieved by turning time-domain samplings to the dual domain via short-time temporal Fourier transformation. The measurement band can be positioned freely within the low radio-frequency (RF) spectrum by tuning the frequency of the reference beam in real-time. Video-rate image rendering is achieved by streamline image processing with commodity computer graphics hardware. This experimental scheme is validated by a non-contact vibrometry experiment.