In vivo bidirectional color Doppler flow imaging of picoliter blood volumes using optical coherence tomography

We describe a novel optical system for bidirectional color Doppler imaging of flow in biological tissues with micrometer-scale resolution and demonstrate its use for in vivo imaging of blood flow in an animal model. Our technique, color Doppler optical coherence tomography (CDOCT), performs spatially localized optical Doppler velocimetry by use of scanning low-coherence interferometry. CDOCT is an extension of optical coherence tomography (OCT), employing coherent signal-acquisition electronics and joint time-frequency analysis algorithms to perform flow imaging simultaneous with conventional OCT imaging. Cross-sectional maps of blood flow velocity with <50-microm spatial resolution and <0.6-mm/s velocity precision were obtained through intact skin in living hamster subdermal tissue. This technology has several potential medical applications.     

High-flow-velocity and shear-rate imaging by use of color Doppler optical coherence tomography

Color Doppler optical coherence tomography (CDOCT) is capable of precise velocity mapping in turbid media. Previous CDOCT systems based on the short-time Fourier transform have been limited to maximum flow velocities of the order of tens of millimeters per second. We describe a technique, based on interference signal demodulation at multiple frequencies, to extend the physiological relevance of CDOCT by increasing the dynamic range of measurable velocities to hundreds of millimeters per second. The physiologically important parameter of shear rate is also derived from CDOCT measurements. The measured flow-velocity profiles and shear-rate distributions correlate very well with theoretical predictions. The multiple demodulation technique, therefore, may be useful to monitor blood flow in vivo and to identify regions with high and low shear rates.     

Real-time, high velocity-resolution color Doppler optical coherence tomography

Color Doppler optical coherence tomography (CDOCT) is a noninvasive optical imaging technique for micrometer-scale physiological flow mapping simultaneously with morphological optical coherence tomography imaging. We have developed a novel CDOCT signal-processing strategy capable of imaging physiological flow rates at 8 frames/s. Our new strategy features hardware-implemented digital autocorrelation across subsequent scans, permitting us to measure 300-Hz-8-kHz Doppler shifts upon signals of 0.6-MHz bandwidth. The performance of the CDOCT system was demonstrated in a flow phantom and in vivo in Xenopus laevis.     

Velocity-estimation accuracy and frame-rate limitations in color Doppler optical coherence tomography

Color Doppler optical coherence tomography (CDOCT) is a recent innovation that allows spatially localized flow-velocity mapping simultaneously with microstructural imaging. We present a theoretical model for velocity-image formation in CDOCT. The proportionality between the heterodyne detector current Doppler power spectrum in CDOCT and the optical source power spectrum is established. We show that stochastic modifications of the Doppler spectrum by fluctuating scatterer distributions in the flow field give rise to unavoidable velocity-estimation inaccuracies as well as to a fundamental trade-off between image-acquisition rate and velocity precision. Novel algorithms that permit high-fidelity depth-resolved measurements of velocities in turbid media are also reported.     

Quantitative retinal-blood flow measurement with three-dimensional vessel geometry determination using ultrahigh-resolution Doppler optical coherence angiography

Retinal blood flow quantification by retinal vessel segmentation with Doppler optical coherence angiography is presented. Vessel diameter, orientation, and position are determined in an en face vessel image and two representative cross-sectional flow images of the vessel. Absolute blood flow velocity is calculated with the help of the measured Doppler frequency shift and determined vessel angle. The volumetric flow rate is obtained with the position and the region of the vessel lumen. The volumetric blood flow rate of retinal arteries before and after a bifurcation is verified in a healthy human eye.     

Measurements of blood flow and blood concentration change using laser speckle in fiber illumination and its application to estimation of stress condition

Optical Review - Speckle imaging method is useful for monitoring of blood flow in living bodies. We have proposed so far the method for simultaneous imaging of blood flow and blood concentration...     

Depth measurement of a blood flow region based on speckle decorrelation

Optical Review - Laser speckle technique is a useful means for imaging blood flow in living bodies and, it has been utilized for analyzing the condition of living bodies or the health state....     

Retinal flow cytometer

The in vivo flow cytometer is an instrument capable of continuous, real-time monitoring of fluorescently labeled cells in the circulation without the need to draw blood samples. However, the original system probes a single vessel in the mouse ear; the small sample volume limits the sensitivity of the technique. We describe an in vivo retinal flow cytometer that simultaneously probes five artery-vein pairs in the mouse eye by circularly scanning a small laser spot rapidly around the optic nerve head. We demonstrate that the retinal flow cytometer detects about five times more cells per minute than the original in vivo flow cytometer does in the ear.     

Simplified laser-speckle-imaging analysis method and its application to retinal blood flow imaging

Laser speckle imaging (LSI) is widely used to study blood flow at high spatiotemporal resolution. Several papers recently pointed out that the commonly used LSI equation involves an approximation that could result in incorrect data analysis. We investigated the impact of such an approximation and introduced a simplified analysis method to improve computation time. Flow phantom studies were performed for validation. Moreover, we demonstrated a novel LSI application by imaging blood flow of rat retinas under normal and physiologic-challenge conditions. Because blood-flow abnormality is implicated in many retinal diseases, LSI could provide valuable physiologic, and potentially diagnostic, information.     

Balanced steady state free precession for arterial spin labeling MRI: Initial experience for blood flow mapping in human brain,retina, and kidney

We implemented pseudo-continuous ASL (pCASL) with 2D and 3D balanced steady state free precession (bSSFP) readout for mapping blood flow in the human brain, retina, and kidney, free of distortion and signal dropout, which are typically observed in the most commonly used echo-planar imaging acquisition. High resolution functional brain imaging in the human visual cortex was feasible with 3D bSSFP pCASL. Blood flow of the human retina could be imaged with pCASL and bSSFP in conjunction with a phase cycling approach to suppress the banding artifacts associated with bSSFP. Furthermore, bSSFP based pCASL enabled us to map renal blood flow within a single breath hold. Control and test–retest experiments suggested that the measured blood flow values in retina and kidney were reliable. Because there is no specific imaging tool for mapping human retina blood flow and the standard contrast agent technique for mapping renal blood flow can cause problems for patients with kidney dysfunction, bSSFP based pCASL may provide a useful tool for the diagnosis of retinal and renal diseases and can complement existing imaging techniques.     

Flow visualization by means of single-exposure speckle photography

A photograph of a flow field is taken under laser illumination. If the exposure is short enough, the velocity distribution in the field will be mapped on the photograph as variations in speckle contrast. These contrast variations can be converted to intensity variations by means of a simple spatial filtering technique, to give a direct picture of the velocity distribution in the flow field. A potential application of this technique to the mapping of retinal blood flow is described.     

Simultaneous blood flow and oxygenation measurements using an off-the-shelf spectrometer

Blood oxygenation and flow are both important parameters in a living body. In this Letter, we introduce a simple configuration to simultaneously measure blood flow and oxygenation using an off-the-shelf spectrometer. With the integration time of 10 ms, flow phantom measurements, a liquid blood phantom test, and an arm cuff occlusion paradigm were performed to validate the feasibility of the system. We expect this proof-of-concept study would be widely adopted by other researchers for acquiring both blood flow and oxygenation changes due to its straightforward configuration and the possibility of multimodal measurement.     

Mechanism and properties of thermally biological effect of Micro waves

水在生命体中是大量存在的,它是维持生命的必要物质.它能促进细胞的繁殖和血液的循环,促进和参与生物化学反应,为生物大分子的功能发挥提供适当条件.由于它是具有一定偶极矩的微观小分子,其转动频率是处在微波范围内,所以我们判定生物体中的液态水能吸收一定频率和强度的微波,并可以将所吸收的微波能量转化为水分子无规运动的热能,使水温升高.我们的实验结果应证了这个机制,从而可导致血液循环,细胞繁殖和一些生化反应的加快和增强DNA,蛋白质和酶的生物活性等生物功能.这就是毫米波的生物热效应的机理和特性.     

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.     

Real-time blind deconvolution of retinal images in adaptive optics scanning laser ophthalmoscopy

With the use of adaptive optics (AO), the ocular aberrations can be compensated to get high-resolution image of living human retina. However, the wavefront correction is not perfect due to the wavefront measure error and hardware restrictions. Thus, it is necessary to use a deconvolution algorithm to recover the retinal images. In this paper, a blind deconvolution technique called Incremental Wiener filter is used to restore the adaptive optics confocal scanning laser ophthalmoscope (AOSLO) images. The point-spread function (PSF) measured by wavefront sensor is only used as an initial value of our algorithm. We also realize the Incremental Wiener filter on graphics processing unit (GPU) in real-time. When the image size is 512 × 480 pixels, six iterations of our algorithm only spend about 10 ms. Retinal blood vessels as well as cells in retinal images are restored by our algorithm, and the PSFs are also revised. Retinal images with and without adaptive optics are both restored. The results show that Incremental Wiener filter reduces the noises and improve the image quality.     

Biophysical aspects of effects of laser radiation on living systems: I. Effect on laboratory animals

It is shown that, taking into account tendencies in the development of new laser technologies, the study of the effect of laser light on humans and animals is a topical and important ecological problem. The results of studies of the effect of coherent (laser) and partially coherent radiations on the organism of white rats are presented. The results obtained for this model can be extrapolated to humans. Biotesting was performed using the methods of high-spatial-resolution Doppler diagnostics and analysis of the contrast of laser speckles. It is found that the time coherence of light does not affect the cerebral blood flow and does not cause reliable changes in the blood microcirculation in the cutaneous covering. It is also reliably shown that neither monochromatic nor quasi-monochromatic laser radiations change the stability of living systems. It is demonstrated that laser sources in new video technologies have no effect on the blood microcirculation in individual organs.     

In vivo monitoring of multiple circulating cell populations using two-photon flow cytometry

To detect and quantify multiple distinct populations of cells circulating simultaneously in the blood of living animals, we developed a novel optical system for two-channel, two-photon flow cytometry in vivo. We used this system to investigate the circulation dynamics in live animals of breast cancer cells with low (MCF-7) and high (MDA-MB-435) metastatic potential, showing for the first time that two different populations of circulating cells can be quantified simultaneously in the vasculature of a single live mouse. We also non-invasively monitored a population of labeled, circulating red blood cells for more than two weeks, demonstrating that this technique can also quantify the dynamics of abundant cells in the vascular system for prolonged periods of time. These data are the first in vivo application of multichannel flow cytometry utilizing two-photon excitation, which will greatly enhance our capability to study circulating cells in cancer and other disease processes.     

Adaptive optics retinal image registration from scale-invariant feature transform

With the use of adaptive optics(AO), high-resolution microscopic imaging of the living human retina in the single cell level has been achieved. In an AO retinal imaging system, with a small field size (about 1°, 300 μm) the motion of the eye severely affects the stabilization of the real-time video images results in significant distortions of the retina images. Scale-invariant feature transform (SIFT) algorithm is applied to automatically abstract corner points with subpixel resolution and match the points in two frames. With the matched corner points, we estimate and remove the motions of 20 frames of photoreceptor cells and capillary blood vessels, respectively. The maximal translational motion is about 30 and 44 pixels in the 20 frames whose size is 416 × 416 pixels. More general motions can be considered by the SIFT algorithm, but only simple translational motion can be considered by cross-correlation algorithm.     

Double-pass imaging polarimetry in the human eye

A Mueller-matrix imaging polarimeter was developed to measure spatially resolved polarization properties in the living human eye. The apparatus is a double-pass setup that incorporates two liquid-crystal variable retarders and a slow-scan CCD camera in the recording stage. Series of 16 images for the combinations of independent polarization states in the first and second passages were recorded for two experimental conditions: with the camera conjugated either with the retina or with the eye's pupil plane. Spatially resolved collections of Mueller matrices and the degree of polarization were calculated from those images for both retinal and pupil planes.     

On the nature of the BOLD fMRI contrast mechanism

Since its development about 15 years ago, functional magnetic resonance imaging (fMRI) has become the leading research tool for mapping brain activity. The technique works by detecting the levels of oxygen in the blood, point by point, throughout the brain. In other words, it relies on a surrogate signal, resulting from changes in oxygenation, blood volume and flow, and does not directly measure neural activity. Although a relationship between changes in brain activity and blood flow has long been speculated, indirectly examined and suggested and surely anticipated and expected, the neural basis of the fMRI signal was only recently demonstrated directly in experiments using combined imaging and intracortical recordings. In the present paper, we discuss the results obtained from such combined experiments. We also discuss our current knowledge of the extracellularly measured signals of the neural processes that they represent and of the structural and functional neurovascular coupling, which links such processes with the hemodynamic changes that offer the surrogate signal that we use to map brain activity. We conclude by considering applications of invasive MRI, including injections of paramagnetic tracers for the study of connectivity in the living animal and simultaneous imaging and electrical microstimulation.