Automated detection of hematological abnormalities through classification of flow cytometric data patterns

Flow Cytometry analyzers have become trusted companions due to their ability to perform fast and accurate analyses of human blood. The aim of these analyses is to determine the possible existence of abnormalities in the blood that have been correlated with serious disease states, such as infectious mononucleosis, leukemia, and various cancers. Though these analyzers provide important feedback, it is always desired to improve the accuracy of the results. This is evidenced by the occurrences of misclassifications reported by some users of these devices. It is advantageous to provide a pattern interpretation framework that is able to provide better classification ability than is currently available. Toward this end, the purpose of this dissertation was to establish a feature extraction and pattern classification framework capable of providing improved accuracy for detecting specific hematological abnormalities in flow cytometric blood data. ^ This involved extracting a unique and powerful set of shift-invariant statistical features from the multi-dimensional flow cytometry data and then using these features as inputs to a pattern classification engine composed of an artificial neural network (ANN). The contribution of this method consisted of developing a descriptor matrix that can be used to reliably assess if a donor’s blood pattern exhibits a clinically abnormal level of variant lymphocytes, which are blood cells that are potentially indicative of disorders such as leukemia and infectious mononucleosis. ^ This study showed that the set of shift-and-rotation-invariant statistical features extracted from the eigensystem of the flow cytometric data pattern performs better than other commonly-used features in this type of disease detection, exhibiting an accuracy of 80.7%, a sensitivity of 72.3%, and a specificity of 89.2%. This performance represents a major improvement for this type of hematological classifier, which has historically been plagued by poor performance, with accuracies as low as 60% in some cases. This research ultimately shows that an improved feature space was developed that can deliver improved performance for the detection of variant lymphocytes in human blood, thus providing significant utility in the realm of suspect flagging algorithms for the detection of blood-related diseases.^

A mathematical resolution to log transformations and the binning effect in applied processing of data in flow cytometry

This dissertation develops a new mathematical approach that overcomes the effect of a data processing phenomenon known as “histogram binning” inherent to flow cytometry data. A real-time procedure is introduced to prove the effectiveness and fast implementation of such an approach on real-world data. The histogram binning effect is a dilemma posed by two seemingly antagonistic developments: (1) flow cytometry data in its histogram form is extended in its dynamic range to improve its analysis and interpretation, and (2) the inevitable dynamic range extension introduces an unwelcome side effect, the binning effect, which skews the statistics of the data, undermining as a consequence the accuracy of the analysis and the eventual interpretation of the data. ^ Researchers in the field contended with such a dilemma for many years, resorting either to hardware approaches that are rather costly with inherent calibration and noise effects; or have developed software techniques based on filtering the binning effect but without successfully preserving the statistical content of the original data. ^ The mathematical approach introduced in this dissertation is so appealing that a patent application has been filed. The contribution of this dissertation is an incremental scientific innovation based on a mathematical framework that will allow researchers in the field of flow cytometry to improve the interpretation of data knowing that its statistical meaning has been faithfully preserved for its optimized analysis. Furthermore, with the same mathematical foundation, proof of the origin of such an inherent artifact is provided. ^ These results are unique in that new mathematical derivations are established to define and solve the critical problem of the binning effect faced at the experimental assessment level, providing a data platform that preserves its statistical content. ^ In addition, a novel method for accumulating the log-transformed data was developed. This new method uses the properties of the transformation of statistical distributions to accumulate the output histogram in a non-integer and multi-channel fashion. Although the mathematics of this new mapping technique seem intricate, the concise nature of the derivations allow for an implementation procedure that lends itself to a real-time implementation using lookup tables, a task that is also introduced in this dissertation. ^

A tidal creek water budget: Estimation of groundwater discharge and overland flow using hydrologic modeling in the Southern Everglades

Taylor Slough is one of the natural freshwater contributors to Florida Bay through a network of microtidal creeks crossing the Everglades Mangrove Ecotone Region (EMER). The EMER ecological function is critical since it mediates freshwater and nutrient inputs and controls the water quality in Eastern Florida Bay. Furthermore, this region is vulnerable to changing hydrodynamics and nutrient loadings as a result of upstream freshwater management practices proposed by the Comprehensive Everglades Restoration Program (CERP), currently the largest wetland restoration project in the USA. Despite the hydrological importance of Taylor Slough in the water budget of Florida Bay, there are no fine scale (∼1 km2) hydrodynamic models of this system that can be utilized as a tool to evaluate potential changes in water flow, salinity, and water quality. Taylor River is one of the major creeks draining Taylor Slough freshwater into Florida Bay. We performed a water budget analysis for the Taylor River area, based on long-term hydrologic data (1999–2007) and supplemented by hydrodynamic modeling using a MIKE FLOOD (DHI,http://dhigroup.com/) model to evaluate groundwater and overland water discharges. The seasonal hydrologic characteristics are very distinctive (average Taylor River wet vs. dry season outflow was 6 to 1 during 1999–2006) with a pronounced interannual variability of flow. The water budget shows a net dominance of through flow in the tidal mixing zone, while local precipitation and evapotranspiration play only a secondary role, at least in the wet season. During the dry season, the tidal flood reaches the upstream boundary of the study area during approximately 80 days per year on average. The groundwater field measurements indicate a mostly upwards-oriented leakage, which possibly equals the evapotranspiration term. The model results suggest a high importance of groundwater contribution to the water salinity in the EMER. The model performance is satisfactory during the dry season where surface flow in the area is confined to the Taylor River channel. The model also provided guidance on the importance of capturing the overland flow component, which enters the area as sheet flow during the rainy season. Overall, the modeling approach is suitable to reach better understanding of the water budget in the mangrove region. However, more detailed field data is needed to ascertain model predictions by further calibrating overland flow parameters.

A mathematical resolution to log transformations and the binning effect in applied processing of data in flow cytometry

This dissertation develops a new mathematical approach that overcomes the effect of a data processing phenomenon known as "histogram binning" inherent to flow cytometry data. A real-time procedure is introduced to prove the effectiveness and fast implementation of such an approach on real-world data. The histogram binning effect is a dilemma posed by two seemingly antagonistic developments: (1) flow cytometry data in its histogram form is extended in its dynamic range to improve its analysis and interpretation, and (2) the inevitable dynamic range extension introduces an unwelcome side effect, the binning effect, which skews the statistics of the data, undermining as a consequence the accuracy of the analysis and the eventual interpretation of the data. Researchers in the field contended with such a dilemma for many years, resorting either to hardware approaches that are rather costly with inherent calibration and noise effects; or have developed software techniques based on filtering the binning effect but without successfully preserving the statistical content of the original data. The mathematical approach introduced in this dissertation is so appealing that a patent application has been filed. The contribution of this dissertation is an incremental scientific innovation based on a mathematical framework that will allow researchers in the field of flow cytometry to improve the interpretation of data knowing that its statistical meaning has been faithfully preserved for its optimized analysis. Furthermore, with the same mathematical foundation, proof of the origin of such an inherent artifact is provided. These results are unique in that new mathematical derivations are established to define and solve the critical problem of the binning effect faced at the experimental assessment level, providing a data platform that preserves its statistical content. In addition, a novel method for accumulating the log-transformed data was developed. This new method uses the properties of the transformation of statistical distributions to accumulate the output histogram in a non-integer and multi-channel fashion. Although the mathematics of this new mapping technique seem intricate, the concise nature of the derivations allow for an implementation procedure that lends itself to a real-time implementation using lookup tables, a task that is also introduced in this dissertation.