Imaging White Blood Cells using a Snapshot Hyper-Spectral Imaging System

Automated white blood cell (WBC) counting systems process an extracted whole blood sample and provide a cell count. A step that would not be ideal for onsite screening of individuals in triage or at a security gate. Snapshot Hyper-Spectral imaging systems are capable of capturing several spectral bands simultaneously, offering co-registered images of a target. With appropriate optics, these systems are potentially able to image blood cells in vivo as they flow through a vessel, eliminating the need for a blood draw and sample staining. Our group has evaluated the capability of a commercial Snapshot Hyper-Spectral imaging system, specifically the Arrow system from Rebellion Photonics, in differentiating between white and red blood cells on unstained and sealed blood smear slides. We evaluated the imaging capabilities of this hyperspectral camera as a platform to build an automated blood cell counting system. Hyperspectral data consisting of 25, 443x313 hyperspectral bands with ~3nm spacing were captured over the range of 419 to 494nm. Open-source hyperspectral datacube analysis tools, used primarily in Geographic Information Systems (GIS) applications, indicate that white blood cells\u27 features are most prominent in the 428-442nm band for blood samples viewed under 20x and 50x magnification over a varying range of illumination intensities. The system has shown to successfully segment blood cells based on their spectral-spatial information. These images could potentially be used in subsequent automated white blood cell segmentation and counting algorithms for performing in vivo white blood cell counting

Blood Cell Revolution: Unveiling 11 Distinct Types with ‘Naturalize’ Augmentation

Artificial intelligence (AI) has emerged as a cutting-edge tool, simultaneously accelerating, securing, and enhancing the diagnosis and treatment of patients. An exemplification of this capability is evident in the analysis of peripheral blood smears (PBS). In university medical centers, hematologists routinely examine hundreds of PBS slides daily to validate or correct outcomes produced by advanced hematology analyzers assessing samples from potentially problematic patients. This process may logically lead to erroneous PBC readings, posing risks to patient health. AI functions as a transformative tool, significantly improving the accuracy and precision of readings and diagnoses. This study reshapes the parameters of blood cell classification, harnessing the capabilities of AI and broadening the scope from 5 to 11 specific blood cell categories with the challenging 11-class PBC dataset. This transformation facilitates a more profound exploration of blood cell diversity, surpassing prior constraints in medical image analysis. Our approach combines state-of-the-art deep learning techniques, including pre-trained ConvNets, ViTb16 models, and custom CNN architectures. We employ transfer learning, fine-tuning, and ensemble strategies, such as CBAM and Averaging ensembles, to achieve unprecedented accuracy and interpretability. Our fully fine-tuned EfficientNetV2 B0 model sets a new standard, with a macro-average precision, recall, and F1-score of 91%, 90%, and 90%, respectively, and an average accuracy of 93%. This breakthrough underscores the transformative potential of 11-class blood cell classification for more precise medical diagnoses. Moreover, our groundbreaking “Naturalize” augmentation technique produces remarkable results. The 2K-PBC dataset generated with “Naturalize” boasts a macro-average precision, recall, and F1-score of 97%, along with an average accuracy of 96% when leveraging the fully fine-tuned EfficientNetV2 B0 model. This innovation not only elevates classification performance but also addresses data scarcity and bias in medical deep learning. Our research marks a paradigm shift in blood cell classification, enabling more nuanced and insightful medical analyses. The “Naturalize” technique’s impact extends beyond blood cell classification, emphasizing the vital role of diverse and comprehensive datasets in advancing healthcare applications through deep learning.This work is supported by grant PID2021-126701OB-I00 funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe”, and by grant GIU19/027 funded by the University of the Basque Country UPV/EHU

Trainable COSFIRE filters for vessel delineation with application to retinal images

Retinal imaging provides a non-invasive opportunity for the diagnosis of several medical pathologies. The automatic segmentation of the vessel tree is an important pre-processing step which facilitates subsequent automatic processes that contribute to such diagnosis. We introduce a novel method for the automatic segmentation of vessel trees in retinal fundus images. We propose a filter that selectively responds to vessels and that we call B-COSFIRE with B standing for bar which is an abstraction for a vessel. It is based on the existing COSFIRE (Combination Of Shifted Filter Responses) approach. A B-COSFIRE filter achieves orientation selectivity by computing the weighted geometric mean of the output of a pool of Difference-of-Gaussians filters, whose supports are aligned in a collinear manner. It achieves rotation invariance efficiently by simple shifting operations. The proposed filter is versatile as its selectivity is determined from any given vessel-like prototype pattern in an automatic configuration process. We configure two B-COSFIRE filters, namely symmetric and asymmetric, that are selective for bars and bar-endings, respectively. We achieve vessel segmentation by summing up the responses of the two rotation-invariant B-COSFIRE filters followed by thresholding. The results that we achieve on three publicly available data sets (DRIVE: Se = 0.7655, Sp = 0.9704; STARE: Se = 0.7716, Sp = 0.9701; CHASE_DB1: Se = 0.7585, Sp = 0.9587) are higher than many of the state-of-the-art methods. The proposed segmentation approach is also very efficient with a time complexity that is significantly lower than existing methods.peer-reviewe

Methods for Analysing Endothelial Cell Shape and Behaviour in Relation to the Focal Nature of Atherosclerosis

The aim of this thesis is to develop automated methods for the analysis of the spatial patterns, and the functional behaviour of endothelial cells, viewed under microscopy, with applications to the understanding of atherosclerosis. Initially, a radial search approach to segmentation was attempted in order to trace the cell and nuclei boundaries using a maximum likelihood algorithm; it was found inadequate to detect the weak cell boundaries present in the available data. A parametric cell shape model was then introduced to fit an equivalent ellipse to the cell boundary by matching phase-invariant orientation fields of the image and a candidate cell shape. This approach succeeded on good quality images, but failed on images with weak cell boundaries. Finally, a support vector machines based method, relying on a rich set of visual features, and a small but high quality training dataset, was found to work well on large numbers of cells even in the presence of strong intensity variations and imaging noise. Using the segmentation results, several standard shear-stress dependent parameters of cell morphology were studied, and evidence for similar behaviour in some cell shape parameters was obtained in in-vivo cells and their nuclei. Nuclear and cell orientations around immature and mature aortas were broadly similar, suggesting that the pattern of flow direction near the wall stayed approximately constant with age. The relation was less strong for the cell and nuclear length-to-width ratios. Two novel shape analysis approaches were attempted to find other properties of cell shape which could be used to annotate or characterise patterns, since a wide variability in cell and nuclear shapes was observed which did not appear to fit the standard parameterisations. Although no firm conclusions can yet be drawn, the work lays the foundation for future studies of cell morphology. To draw inferences about patterns in the functional response of cells to flow, which may play a role in the progression of disease, single-cell analysis was performed using calcium sensitive florescence probes. Calcium transient rates were found to change with flow, but more importantly, local patterns of synchronisation in multi-cellular groups were discernable and appear to change with flow. The patterns suggest a new functional mechanism in flow-mediation of cell-cell calcium signalling

Automated Detection of Acute Leukemia using K-mean Clustering Algorithm

Leukemia is a hematologic cancer which develops in blood tissue and triggers rapid production of immature and abnormal shaped white blood cells. Based on statistics it is found that the leukemia is one of the leading causes of death in men and women alike. Microscopic examination of blood sample or bone marrow smear is the most effective technique for diagnosis of leukemia. Pathologists analyze microscopic samples to make diagnostic assessments on the basis of characteristic cell features. Recently, computerized methods for cancer detection have been explored towards minimizing human intervention and providing accurate clinical information. This paper presents an algorithm for automated image based acute leukemia detection systems. The method implemented uses basic enhancement, morphology, filtering and segmenting technique to extract region of interest using k-means clustering algorithm. The proposed algorithm achieved an accuracy of 92.8% and is tested with Nearest Neighbor (KNN) and Naive Bayes Classifier on the data-set of 60 samples.Comment: Presented in ICCCCS 201

DeepSegNet: An Innovative Framework for Accurate Blood Cell Image Segmentation

Image segmentation plays a crucial and indispensable role in computer vision, as it allows the partitioning of an image into meaningful regions or objects. Among its numerous applications, image segmentation holds particular significance in the domains of medical diagnosis and healthcare. Its vital role in this field stems from its ability to extract and delineate specific anatomical structures, tumors, lesions, and other critical regions from medical images. In medical diagnosis, accurate and precise segmentation of organs and abnormalities is paramount for effective treatment planning, disease monitoring, and surgical interventions. Blood cell image segmentation is highly valuable for medical diagnosis and research, particularly in the domains of hematology and pathology. Precisely segmenting blood cells from microscopic images is essential, as it offers critical insights into various blood-related disorders and diseases. Although deep learning segmentation models have exhibited promising results in blood cell image segmentation, they suffer from several limitations. These drawbacks encompass scarce data availability, inefficient feature extraction, extended computation time, limited generalization to unseen data, challenges with variations, and artifacts. Consequently, these limitations can adversely impact the overall performance of the models. Blood cell image segmentation encounters persistent challenges due to factors like irregular cell shapes, which pose difficulties in boundary delineation, imperfect cell separation in smears, and low cell contrast, leading to visibility issues during segmentation. This research article introduces the innovative DeepSegNet framework, a powerful solution for precise blood cell image segmentation. The performance of widely-used segmentation models like PSPNet, FPN, and DeepLabv3+ is enhanced through the use of sophisticated preprocessing techniques, improving generalization capability, data diversity, and training stability. Additionally, the incorporation of diverse dilated convolutions and feature fusion further contributes to the improvement of these models. The Improved PSPNet, Improved FPN, Deep Lab V3, and Improved Deep Lab V3+ achieved 98.25%, 99.04%, 98.23%, and 99.31% accuracy, respectively, and the Improved Deep Lab V3+ model outperformed well and produced a Dice Coefficient of 99.32% and Precision of 99.38%. The proposed DeepSegNet framework improves overall performance with an increased accuracy of 8.91%, 3.72%, 17.73%, 22.83%, 7.96%, 9.61%, 17.36%, 6.22%, 13.32%, and 14.32% compared to the existing models. This framework, which can be applied to accurately identify and quantify different cell types from blood cell images, is instrumental in diagnosing a variety of hematological disorders and diseases

Computer vision for microscopy diagnosis of malaria

This paper reviews computer vision and image analysis studies aiming at automated diagnosis or screening of malaria infection in microscope images of thin blood film smears. Existing works interpret the diagnosis problem differently or propose partial solutions to the problem. A critique of these works is furnished. In addition, a general pattern recognition framework to perform diagnosis, which includes image acquisition, pre-processing, segmentation, and pattern classification components, is described. The open problems are addressed and a perspective of the future work for realization of automated microscopy diagnosis of malaria is provided