Dingan Han

In vivo label-free microangiography by laser speckle imaging with intensity fluctuation modulation

Abstract. We present the theory of laser speckle imaging improved with intensity fluctuation modulation, where the dynamic speckle pattern can be isolated from its stationary counterpart. A series of in vivo experiments demonstrate the effectiveness of our method in achieving microangiography and monitoring vascular self-recovering process. All results show the convincing performance of our imaging method in both structural and functional imaging of blood flow, which may have potential applications in biological research and disease diagnosis.

Full-field optical micro-angiography

We present a detailed description of full-field optical micro-angiography on the basis of frequency-domain laser speckle imaging with intensity fluctuation modulation (LSI-IFM). The imaging approach works based on the instantaneous local intensity fluctuation realized via the combination of short exposure and low sampling rate of a camera and appropriate magnification of a microscope. In vivo experiments on mouse ear verify the theoretical description we made for the imaging mechanism and demonstrate the ability of LSI-IFM as optical micro-angiography. By introducing a fundus camera into LSI-IFM system, our approach has a potential application in label-free retina optical micro-angiography.

Laser speckle projection tomography

We propose a laser speckle projection tomography (LSPT) method to obtain a three-dimensional (3D) flowing image. The method combines the advantages of optical projection tomography and laser speckle imaging to reconstruct the visualization of 3D flowing structure. With LSPT, the flowing signal is extracted by laser speckle contrast method and the 3D flowing image is reconstructed by the filtered back-projection algorithm. A phantom experiment is performed to demonstrate that LSPT is able to obtain 3D flowing structure, influenced by concentration and the flow speed.

Laser Doppler projection tomography

We propose a laser Doppler projection tomography (LDPT) method to obtain visualization of three-dimensional (3D) flowing structures. With LDPT, the flowing signal is extracted by a modified laser Doppler method, and the 3D flowing image is reconstructed by the filtered backprojection algorithm. Phantom experiments are performed to demonstrate that LDPT is able to obtain 3D flowing structure with higher signal-to-noise ratio and spatial resolution. Our experiment results display its potentially useful application to develop 3D label-free optical angiography for the circulation system of live small animal models or microfluidic experiments.

In vivo full-field functional optical hemocytometer

We present an in vivo lab-free full-field functional optical hemocytometer (FFOH) for application to the capillaries of a live biological specimen, based on the absorption intensity fluctuation modulation (AIFM) effect. Because of the absorption difference between the red blood cells (RBCs) and background tissue under low-coherence light illumination, an endogenous instantaneous intensity fluctuation is generated by the AIFM effect when RBCs discontinuously traverse the capillary. The AIFM effect is used to highlight the RBC signal relative to the background tissue by computing the real-time modulation depth. FFOH can simultaneously provide a flow video, the flow velocity and the RBC count. Ourexperimental results can potentially be applied to study the physiological mechanisms of the blood circulation systems of near-transparent live biological samples.

Large-depth-of-field full-field optical angiography

A large-depth-of-field full-field optical angiography (LD-FFOA) method is developed to expand the depth-of-field (DOF) using a contrast pyramid fusion algorithm (CPFA). The absorption intensity fluctuation modulation effect is utilized to obtain full-field optical angiography (FFOA) images at different focus positions. The CPFA is used to process these FFOA images with different focuses. By selecting high-contrast areas, the CPFA can highlight the characteristics and details of blood vessels to obtain LD-FFOA images. In the optimal case of the proposed method, the DOF for FFOA is more than tripled using 10 differently focused FFOA images. Both the phantom and animal experimental results show that the LD-FFOA resolves FFOA defocusing issues induced by surface and thickness inhomogeneities in biological samples. The proposed method can be potentially applied to practical biological experiments.

Full-field functional optical angiography

We propose full-field functional optical angiography for a live biological specimen based on the absorption intensity fluctuation modulation (AIFM) effect. Because of the difference in absorption between red blood cells (RBCs) and the background tissue under low-coherence light illumination, the moving RBCs, which discontinuously pass though the capillary vessels, generate an AIFM effect. This effect offers high contrast of absorption imaging and sensitivity of low-coherence interference between RBCs and the background tissue. It is used to distinguish the signal of RBCs from that of the background tissue. The averaged and real-time modulation depths are computed to obtain full-field label-free optical angiography and measure blood flow velocity simultaneously. The AIFM method could potentially be applied to study the physiological mechanisms of blood circulation systems of near-transparent live biologic samples.

High-temporal-resolution, full-field optical angiography based on short-time modulation depth for vascular occlusion tests

Abstract. We developed high-temporal-resolution, full-field optical angiography for use in vascular occlusion tests (VOTs). In the proposed method, undersampled signals are acquired by a high-speed digital camera that separates the dynamic and static speckle signals. The two types of speckle signal are used to calculate the short-time modulation depth (STMD) of each of the camera pixels. STMD is then used to realize high-temporal-resolution, full-field optical angiography. Phantom and biological experiments conducted and demonstrated the feasibility of using our proposed method to perform VOTs and to study the reaction kinetics in microfluidic systems.

Optical projection angiography

We propose the optical projection angiography (OPA) based on lateral dynamic scattering light for visualizing a three-dimensional (3D) blood-flow network. In OPA, a pulsed laser source illuminates a live biological sample for eliminating digital camera integration effects. The 2D flow image can be obtained by separating the dynamic and static scattering light signal of each camera pixel in the frequency domain. Flow images at a different angle are combined to reconstruct the 3D volume of the sample to realize OPA. Moreover, as our experiment retains the bright-field optical projection tomography (OPT) setup, the OPA image for the circulatory system and the OPT image for the skeletal structure can simultaneously be reconstructed. The experimental results can potentially be applied in physiological development studies.

Laser Speckle Micro-angiography

We introduce a laser speckle micro-angiography (LSMA) to reconstruct the 2D visualization of the blood flows. In vivo Biological experiments on the ears of rabbits and mice verify the outstanding performance of our LSMA in imaging microcirculatory network.Provided an potential to develop a label-free optical micro-angiography in retinal imaging.