Visualization and Correction of Automated Segmentation, Tracking and Lineaging from 5-D Stem Cell Image Sequences

Results: We present an application that enables the quantitative analysis of multichannel 5-D (x, y, z, t, channel) and large montage confocal fluorescence microscopy images. The image sequences show stem cells together with blood vessels, enabling quantification of the dynamic behaviors of stem cells in relation to their vascular niche, with applications in developmental and cancer biology. Our application automatically segments, tracks, and lineages the image sequence data and then allows the user to view and edit the results of automated algorithms in a stereoscopic 3-D window while simultaneously viewing the stem cell lineage tree in a 2-D window. Using the GPU to store and render the image sequence data enables a hybrid computational approach. An inference-based approach utilizing user-provided edits to automatically correct related mistakes executes interactively on the system CPU while the GPU handles 3-D visualization tasks. Conclusions: By exploiting commodity computer gaming hardware, we have developed an application that can be run in the laboratory to facilitate rapid iteration through biological experiments. There is a pressing need for visualization and analysis tools for 5-D live cell image data. We combine accurate unsupervised processes with an intuitive visualization of the results. Our validation interface allows for each data set to be corrected to 100% accuracy, ensuring that downstream data analysis is accurate and verifiable. Our tool is the first to combine all of these aspects, leveraging the synergies obtained by utilizing validation information from stereo visualization to improve the low level image processing tasks.Comment: BioVis 2014 conferenc

Multi-Scale 3D Scene Flow from Binocular Stereo Sequences

Scene flow methods estimate the three-dimensional motion field for points in the world, using multi-camera video data. Such methods combine multi-view reconstruction with motion estimation. This paper describes an alternative formulation for dense scene flow estimation that provides reliable results using only two cameras by fusing stereo and optical flow estimation into a single coherent framework. Internally, the proposed algorithm generates probability distributions for optical flow and disparity. Taking into account the uncertainty in the intermediate stages allows for more reliable estimation of the 3D scene flow than previous methods allow. To handle the aperture problems inherent in the estimation of optical flow and disparity, a multi-scale method along with a novel region-based technique is used within a regularized solution. This combined approach both preserves discontinuities and prevents over-regularization – two problems commonly associated with the basic multi-scale approaches. Experiments with synthetic and real test data demonstrate the strength of the proposed approach.National Science Foundation (CNS-0202067, IIS-0208876); Office of Naval Research (N00014-03-1-0108

3D SEM Surface Reconstruction: An Optimized, Adaptive, and Intelligent Approach

Structural analysis of microscopic objects is a longstanding topic in several scientific disciplines, including biological, mechanical, and material sciences. The scanning electron microscope (SEM), as a promising imaging equipment has been around to determine the surface properties (e.g., compositions or geometries) of specimens by achieving increased magnification, contrast, and resolution greater than one nanometer. Whereas SEM micrographs still remain two-dimensional (2D), many research and educational questions truly require knowledge and information about their three-dimensional (3D) surface structures. Having 3D surfaces from SEM images would provide true anatomic shapes of micro samples which would allow for quantitative measurements and informative visualization of the systems being investigated. In this research project, we novel design and develop an optimized, adaptive, and intelligent multi-view approach named 3DSEM++ for 3D surface reconstruction of SEM images, making a 3D SEM dataset publicly and freely available to the research community. The work is expected to stimulate more interest and draw attention from the computer vision and multimedia communities to the fast-growing SEM application area

Light field image processing: an overview

Light field imaging has emerged as a technology allowing to capture richer visual information from our world. As opposed to traditional photography, which captures a 2D projection of the light in the scene integrating the angular domain, light fields collect radiance from rays in all directions, demultiplexing the angular information lost in conventional photography. On the one hand, this higher dimensional representation of visual data offers powerful capabilities for scene understanding, and substantially improves the performance of traditional computer vision problems such as depth sensing, post-capture refocusing, segmentation, video stabilization, material classification, etc. On the other hand, the high-dimensionality of light fields also brings up new challenges in terms of data capture, data compression, content editing, and display. Taking these two elements together, research in light field image processing has become increasingly popular in the computer vision, computer graphics, and signal processing communities. In this paper, we present a comprehensive overview and discussion of research in this field over the past 20 years. We focus on all aspects of light field image processing, including basic light field representation and theory, acquisition, super-resolution, depth estimation, compression, editing, processing algorithms for light field display, and computer vision applications of light field data

Spectrally encoded fiber-based structured lighting probe for intraoperative 3D imaging

Three dimensional quantification of organ shape and structure during minimally invasive surgery (MIS) could enhance precision by allowing the registration of multi-modal or pre-operative image data (US/MRI/CT) with the live optical image. Structured illumination is one technique to obtain 3D information through the projection of a known pattern onto the tissue, although currently these systems tend to be used only for macroscopic imaging or open procedures rather than in endoscopy. To account for occlusions, where a projected feature may be hidden from view and/or confused with a neighboring point, a flexible multispectral structured illumination probe has been developed that labels each projected point with a specific wavelength using a supercontinuum laser. When imaged by a standard endoscope camera they can then be segmented using their RGB values, and their 3D coordinates calculated after camera calibration. The probe itself is sufficiently small (1.7 mm diameter) to allow it to be used in the biopsy channel of commonly used medical endoscopes. Surgical robots could therefore also employ this technology to solve navigation and visualization problems in MIS, and help to develop advanced surgical procedures such as natural orifice translumenal endoscopic surgery

Study of Computational Image Matching Techniques: Improving Our View of Biomedical Image Data

Image matching techniques are proven to be necessary in various fields of science and engineering, with many new methods and applications introduced over the years. In this PhD thesis, several computational image matching methods are introduced and investigated for improving the analysis of various biomedical image data. These improvements include the use of matching techniques for enhancing visualization of cross-sectional imaging modalities such as Computed Tomography (CT) and Magnetic Resonance Imaging (MRI), denoising of retinal Optical Coherence Tomography (OCT), and high quality 3D reconstruction of surfaces from Scanning Electron Microscope (SEM) images. This work greatly improves the process of data interpretation of image data with far reaching consequences for basic sciences research. The thesis starts with a general notion of the problem of image matching followed by an overview of the topics covered in the thesis. This is followed by introduction and investigation of several applications of image matching/registration in biomdecial image processing: a) registration-based slice interpolation, b) fast mesh-based deformable image registration and c) use of simultaneous rigid registration and Robust Principal Component Analysis (RPCA) for speckle noise reduction of retinal OCT images. Moving towards a different notion of image matching/correspondence, the problem of view synthesis and 3D reconstruction, with a focus on 3D reconstruction of microscopic samples from 2D images captured by SEM, is considered next. Starting from sparse feature-based matching techniques, an extensive analysis is provided for using several well-known feature detector/descriptor techniques, namely ORB, BRIEF, SURF and SIFT, for the problem of multi-view 3D reconstruction. This chapter contains qualitative and quantitative comparisons in order to reveal the shortcomings of the sparse feature-based techniques. This is followed by introduction of a novel framework using sparse-dense matching/correspondence for high quality 3D reconstruction of SEM images. As will be shown, the proposed framework results in better reconstructions when compared with state-of-the-art sparse-feature based techniques. Even though the proposed framework produces satisfactory results, there is room for improvements. These improvements become more necessary when dealing with higher complexity microscopic samples imaged by SEM as well as in cases with large displacements between corresponding points in micrographs. Therefore, based on the proposed framework, a new approach is proposed for high quality 3D reconstruction of microscopic samples. While in case of having simpler microscopic samples the performance of the two proposed techniques are comparable, the new technique results in more truthful reconstruction of highly complex samples. The thesis is concluded with an overview of the thesis and also pointers regarding future directions of the research using both multi-view and photometric techniques for 3D reconstruction of SEM images

Development of a handheld fiber-optic probe-based raman imaging instrumentation: raman chemlighter

Raman systems based on handheld fiber-optic probes offer advantages in terms of smaller sizes and easier access to the measurement sites, which are favorable for biomedical and clinical applications in the complex environment. However, there are several common drawbacks of applying probes for many applications: (1) The fixed working distance requires the user to maintain a certain working distance to acquire higher Raman signals; (2) The single-point-measurement ability restricts realizing a mapping or scanning procedure; (3) Lack of real-time data processing and a straightforward co-registering method to link the Raman information with the respective measurement position. The thesis proposed and experimentally demonstrated various approaches to overcome these drawbacks. A handheld fiber-optic Raman probe with an autofocus unit was presented to overcome the problem arising from using fixed-focus lenses, by using a liquid lens as the objective lens, which allows dynamical adjustment of the focal length of the probe. An implementation of a computer vision-based positional tracking to co-register the regular Raman spectroscopic measurements with the spatial location enables fast recording of a Raman image from a large tissue sample by combining positional tracking of the laser spot through brightfield images. The visualization of the Raman image has been extended to augmented and mixed reality and combined with a 3D reconstruction method and projector-based visualization to offer an intuitive and easily understandable way of presenting the Raman image. All these advances are substantial and highly beneficial to further drive the clinical translation of Raman spectroscopy as potential image-guided instrumentation

Determination of volume characteristics of cells from dynamical microscopic image

The algorithm for the determination of 3D-characteristics of a dynamic biological object based on the recovery of a stereo pair was proposed. Images for stereo pair construction were obtained via a single camera before and after the displacement of the object. The algorithm is based on the example of living one-celled organisms