Guest Editorial to the Special Letters Issue on Emerging Technologies in Multiparameter Biomedical Optical Imaging and Image Analysis

The past two decades have witnessed revolutionary advances in biomedical imaging modalities capable of providing biological and physiological information from the cellular scale to the organ level. Recent advances have also been focused on cost-effective, noninvasive, portable, and molecularimaging technologies for imaging at microscopic, mesoscopic, and macroscopic levels. These technologies have significant potential to advance biomedical research and clinical practice. They can also provide a better understanding and monitoring of physiological and functional disorders, which could lead to mainstream diagnostic technologies of the future

Guest Editorial to the Special Letters Issue on Emerging Technologies in Multiparameter Biomedical Optical Imaging and Image Analysis

The past two decades have witnessed revolutionary advances in biomedical imaging modalities capable of providing biological and physiological information from the cellular scale to the organ level. Recent advances have also been focused on cost-effective, noninvasive, portable, and molecularimaging technologies for imaging at microscopic, mesoscopic, and macroscopic levels. These technologies have significant potential to advance biomedical research and clinical practice. They can also provide a better understanding and monitoring of physiological and functional disorders, which could lead to mainstream diagnostic technologies of the future

Individual motile CD4+ T cells can participate in efficient multikilling through conjugation to multiple tumor cells

T cells genetically modified to express a CD19-specific chimeric antigen receptor (CAR) for the investigational treatment of B-cell malignancies comprise a heterogeneous population, and their ability to persist and participate in serial killing of tumor cells is a predictor of therapeutic success. We implemented Timelapse Imaging Microscopy in Nanowell Grids (TIMING) to provide direct evidence that CD4+CAR+ T cells (CAR4 cells) can engage in multikilling via simultaneous conjugation to multiple tumor cells. Comparisons of the CAR4 cells and CD8+CAR+ T cells (CAR8 cells) demonstrate that, although CAR4 cells can participate in killing and multikilling, they do so at slower rates, likely due to the lower granzyme B content. Significantly, in both sets of T cells, a minor subpopulation of individual T cells identified by their high motility demonstrated efficient killing of single tumor cells. A comparison of the multikiller and single-killer CAR+ T cells revealed that the propensity and kinetics of T-cell apoptosis were modulated by the number of functional conjugations. T cells underwent rapid apoptosis, and at higher frequencies, when conjugated to single tumor cells in isolation, and this effect was more pronounced on CAR8 cells. Our results suggest that the ability of CAR+ T cells to participate in multikilling should be evaluated in the context of their ability to resist activation-induced cell death. We anticipate that TIMING may be used to rapidly determine the potency of T-cell populations and may facilitate the design and manufacture of next-generation CAR+ T cells with improved efficacy. Cancer Immunol Res; 3(5); 473–82. ©2015 AACR

An Active Learning Approach for Rapid Characterization of Endothelial Cells in Human Tumors

Currently, no available pathological or molecular measures of tumor angiogenesis predict response to antiangiogenic therapies used in clinical practice. Recognizing that tumor endothelial cells (EC) and EC activation and survival signaling are the direct targets of these therapies, we sought to develop an automated platform for quantifying activity of critical signaling pathways and other biological events in EC of patient tumors by histopathology. Computer image analysis of EC in highly heterogeneous human tumors by a statistical classifier trained using examples selected by human experts performed poorly due to subjectivity and selection bias. We hypothesized that the analysis can be optimized by a more active process to aid experts in identifying informative training examples. To test this hypothesis, we incorporated a novel active learning (AL) algorithm into FARSIGHT image analysis software that aids the expert by seeking out informative examples for the operator to label. The resulting FARSIGHT-AL system identified EC with specificity and sensitivity consistently greater than 0.9 and outperformed traditional supervised classification algorithms. The system modeled individual operator preferences and generated reproducible results. Using the results of EC classification, we also quantified proliferation (Ki67) and activity in important signal transduction pathways (MAP kinase, STAT3) in immunostained human clear cell renal cell carcinoma and other tumors. FARSIGHT-AL enables characterization of EC in conventionally preserved human tumors in a more automated process suitable for testing and validating in clinical trials. The results of our study support a unique opportunity for quantifying angiogenesis in a manner that can now be tested for its ability to identify novel predictive and response biomarkers

Computational prediction of neural progenitor cell fates

Understanding how stem and progenitor cells choose between alternative cell fates is a major challenge in developmental biology. Efforts to tackle this problem have been hampered by the scarcity of markers that can be used to predict cell division outcomes. Here we present a computational method, based on algorithmic information theory, to analyze dynamic features of living cells over time. Using this method, we asked whether rat retinal progenitor cells (RPCs) display characteristic phenotypes before undergoing mitosis that could foretell their fate. We predicted whether RPCs will undergo a self-renewing or terminal division with 99% accuracy, or whether they will produce two photoreceptors or another combination of offspring with 87% accuracy. Our implementation can segment, track and generate predictions for 40 cells simultaneously on a standard computer at 5 min per frame. This method could be used to isolate cell populations with specific developmental potential, enabling previously impossible investigations.The computational aspects of this work were supported by the Center for Subsurface Sensing and Imaging Systems (NSF Grant EEC-9986821), by the Rensselaer Polytechnic Institute and by the University of Wisconsin-Milwaukee. This work was supported by grants from the Canadian Institutes of Health Research and the Foundation Fighting Blindness – Canada (to M.C). M.C. is a CIHR New Investigator and a W.K. Stell Scholar of the Foundation Fighting Blindness – Canada

NetMets: software for quantifying and visualizing errors in biological network segmentation

One of the major goals in biomedical image processing is accurate segmentation of networks embedded in volumetric data sets. Biological networks are composed of a meshwork of thin filaments that span large volumes of tissue. Examples of these structures include neurons and microvasculature, which can take the form of both hierarchical trees and fully connected networks, depending on the imaging modality and resolution. Network function depends on both the geometric structure and connectivity. Therefore, there is considerable demand for algorithms that segment biological networks embedded in three-dimensional data. While a large number of tracking and segmentation algorithms have been published, most of these do not generalize well across data sets. One of the major reasons for the lack of general-purpose algorithms is the limited availability of metrics that can be used to quantitatively compare their effectiveness against a pre-constructed ground-truth. In this paper, we propose a robust metric for measuring and visualizing the differences between network models. Our algorithm takes into account both geometry and connectivity to measure network similarity. These metrics are then mapped back onto an explicit model for visualization

Model-based method for improving the accuracy and repeatability of estimating vascular bifurcations and crossovers from retinal fundus images

A model-based algorithm, termed exclusion region and position refinement (ERPR), is presented for improving the accuracy and repeatability of estimating the locations where vascular structures branch and cross over, in the context of human retinal images. The goal is two fold. First, accurate morphometry of branching and crossover points (landmarks) in neuronal/vascular structure is important to several areas of biology and medicine. Second, these points are valuable as landmarks for image registration, so improved accuracy and repeatability in estimating their locations and signatures leads to more reliable image registration for applications such as change detection and mosaicing. The ERPR algorithm is shown to reduce the median location error from 2.04 pixels down to 1.1 pixels, while improving the median spread (a measure of repeatability) from 2.09 pixels down to 1.05 pixels. Errors in estimating vessel orientations were similarly reduced from 7.2 degrees down to 3.8 degrees

The Embryonic Cell Lineage of the Nematode Caenorhabditis elegans

Abstract This work presents automatic methods for analyzing timelapse 3D confocal image sequences of fluorescently tagged C.elegans worm embryos to extract the lineage, as well as 3D spatial migration paths of cells. Significance Cell lineage of C. elegans is a family-tree representation of cell division pattern, ancestor-descendant relationship, and marker events (e.g. cell death) timeline in embryogenesis. It is essential to genetic, behavioral, and structural analysis of multi-cellular organism development Challenge • Automation with tolerable error rate • User-friendly GUI for human expert edition • Multiple cell measurements for practical analysis Result The automated procedure achieved a 97% accuracy (33 out of 34 cell division events were detected correctly) for a lineage of C. elegans from 56-90 cell stages, covering a span of 140~170mins post fertilization. Technology Transfer Our automated methods can be integrated into a common MHT core for a diversity of problems, e.g. changing detection, multiple objects tracking, etc