Simultaneous Segmentation of Leukocyte and Erythrocyte in Microscopic Images Using a Marker-Controlled Watershed Algorithm

The density or quantity of leukocytes and erythrocytes in a unit volume of blood, which can be automatically measured through a computer-based microscopic image analysis system, is frequently considered an indicator of diseases. The segmentation of blood cells, as a basis of quantitative statistics, plays an important role in the system. However, many conventional methods must firstly distinguish blood cells into two types (i.e., leukocyte and erythrocyte) and segment them in independent procedures. In this paper, we present a marker-controlled watershed algorithm for simultaneously extracting the two types of blood cells to simplify operations and reduce computing time. The method consists of two steps, that is, cell nucleus segmentation and blood cell segmentation. An image enhancement technique is used to obtain the leukocyte marker. Two marker-controlled watershed algorithms are based on distance transformation and edge gradient information to acquire blood cell contour. The segmented leukocytes and erythrocytes are obtained simultaneously by classification. Experimental results demonstrate that the proposed method is fast, robust, and efficient

Improved Otsu and Kapur approach for white blood cells segmentation based on LebTLBO optimization for the detection of Leukemia.

The diagnosis of leukemia involves the detection of the abnormal characteristics of blood cells by a trained pathologist. Currently, this is done manually by observing the morphological characteristics of white blood cells in the microscopic images. Though there are some equipment- based and chemical-based tests available, the use and adaptation of the automated computer vision-based system is still an issue. There are certain software frameworks available in the literature; however, they are still not being adopted commercially. So there is a need for an automated and software- based framework for the detection of leukemia. In software-based detection, segmentation is the first critical stage that outputs the region of interest for further accurate diagnosis. Therefore, this paper explores an efficient and hybrid segmentation that proposes a more efficient and effective system for leukemia diagnosis. A very popular publicly available database, the acute lymphoblastic leukemia image database (ALL-IDB), is used in this research. First, the images are pre-processed and segmentation is done using Multilevel thresholding with Otsu and Kapur methods. To further optimize the segmentation performance, the Learning enthusiasm-based teaching-learning-based optimization (LebTLBO) algorithm is employed. Different metrics are used for measuring the system performance. A comparative analysis of the proposed methodology is done with existing benchmarks methods. The proposed approach has proven to be better than earlier techniques with measuring parameters of PSNR and Similarity index. The result shows a significant improvement in the performance measures with optimizing threshold algorithms and the LebTLBO technique

Threshold estimation based on local minima for nucleus and cytoplasm segmentation

Background Image segmentation is the process of partitioning an image into separate objects or regions. It is an essential step in image processing to segment the regions of interest for further processing. We propose a method for segmenting the nuclei and cytoplasms from white blood cells (WBCs). Methods Initially, the method computes an initial value based on the minimum and maximum values of the input image. Then, a histogram of the input image is computed and approximated to obtain function values. The method searches for the first local maximum and local minimum from the approximated function values in the order of increasing of knots sequence. We approximate the required threshold from the first local minimum and the computed initial value based on defined conditions. The threshold is applied to the input image to binarize it, and then post-processing is performed to obtain the final segmented nucleus. We segment the whole WBC before segmenting the cytoplasm depending on the complexity of the objects in the image. For WBCs that are well separated from red blood cells (RBCs), n thresholds are generated and then produce n thresholded images. Then, a standard Otsu method is used to binarize the average of the produced images. Morphological operations are applied on the binarized image, and then a single-pixel point from the segmented nucleus is used to segment the WBC. For images in which RBCs touch the WBCs, we segment the whole WBC using SLIC and watershed methods. The cytoplasm is obtained by subtracting the segmented nucleus from the segmented WBC. Results The method is tested on two different public data sets and the results are compared to the state of art methods. The performance analysis shows that the proposed method segments the nucleus and cytoplasm well. Conclusion We propose a method for nucleus and cytoplasm segmentation based on the local minima of the approximated function values from the image’s histogram. The method has demonstrated its utility in segmenting nuclei, WBCs, and cytoplasm, and the results are satisfactory.publishedVersio

Robust Method for Semantic Segmentation of Whole-Slide Blood Cell Microscopic Image

Previous works on segmentation of SEM (scanning electron microscope) blood cell image ignore the semantic segmentation approach of whole-slide blood cell segmentation. In the proposed work, we address the problem of whole-slide blood cell segmentation using the semantic segmentation approach. We design a novel convolutional encoder-decoder framework along with VGG-16 as the pixel-level feature extraction model. -e proposed framework comprises 3 main steps: First, all the original images along with manually generated ground truth masks of each blood cell type are passed through the preprocessing stage. In the preprocessing stage, pixel-level labeling, RGB to grayscale conversion of masked image and pixel fusing, and unity mask generation are performed. After that, VGG16 is loaded into the system, which acts as a pretrained pixel-level feature extraction model. In the third step, the training process is initiated on the proposed model. We have evaluated our network performance on three evaluation metrics. We obtained outstanding results with respect to classwise, as well as global and mean accuracies. Our system achieved classwise accuracies of 97.45%, 93.34%, and 85.11% for RBCs, WBCs, and platelets, respectively, while global and mean accuracies remain 97.18% and 91.96%, respectively.Comment: 13 pages, 13 figure

Automatic Segmentation and Classification of Red and White Blood cells in Thin Blood Smear Slides

In this work we develop a system for automatic detection and classification of cytological images which plays an increasing important role in medical diagnosis. A primary aim of this work is the accurate segmentation of cytological images of blood smears and subsequent feature extraction, along with studying related classification problems such as the identification and counting of peripheral blood smear particles, and classification of white blood cell into types five. Our proposed approach benefits from powerful image processing techniques to perform complete blood count (CBC) without human intervention. The general framework in this blood smear analysis research is as follows. Firstly, a digital blood smear image is de-noised using optimized Bayesian non-local means filter to design a dependable cell counting system that may be used under different image capture conditions. Then an edge preservation technique with Kuwahara filter is used to recover degraded and blurred white blood cell boundaries in blood smear images while reducing the residual negative effect of noise in images. After denoising and edge enhancement, the next step is binarization using combination of Otsu and Niblack to separate the cells and stained background. Cells separation and counting is achieved by granulometry, advanced active contours without edges, and morphological operators with watershed algorithm. Following this is the recognition of different types of white blood cells (WBCs), and also red blood cells (RBCs) segmentation. Using three main types of features: shape, intensity, and texture invariant features in combination with a variety of classifiers is next step. The following features are used in this work: intensity histogram features, invariant moments, the relative area, co-occurrence and run-length matrices, dual tree complex wavelet transform features, Haralick and Tamura features. Next, different statistical approaches involving correlation, distribution and redundancy are used to measure of the dependency between a set of features and to select feature variables on the white blood cell classification. A global sensitivity analysis with random sampling-high dimensional model representation (RS-HDMR) which can deal with independent and dependent input feature variables is used to assess dominate discriminatory power and the reliability of feature which leads to an efficient feature selection. These feature selection results are compared in experiments with branch and bound method and with sequential forward selection (SFS), respectively. This work examines support vector machine (SVM) and Convolutional Neural Networks (LeNet5) in connection with white blood cell classification. Finally, white blood cell classification system is validated in experiments conducted on cytological images of normal poor quality blood smears. These experimental results are also assessed with ground truth manually obtained from medical experts

Medical Image Segmentation: Thresholding and Minimum Spanning Trees

I bildesegmentering deles et bilde i separate objekter eller regioner. Det er et essensielt skritt i bildebehandling for å definere interesseområder for videre behandling eller analyse. Oppdelingsprosessen reduserer kompleksiteten til et bilde for å forenkle analysen av attributtene oppnådd etter segmentering. Det forandrer representasjonen av informasjonen i det opprinnelige bildet og presenterer pikslene på en måte som er mer meningsfull og lettere å forstå. Bildesegmentering har forskjellige anvendelser. For medisinske bilder tar segmenteringsprosessen sikte på å trekke ut bildedatasettet for å identifisere områder av anatomien som er relevante for en bestemt studie eller diagnose av pasienten. For eksempel kan man lokalisere berørte eller anormale deler av kroppen. Segmentering av oppfølgingsdata og baseline lesjonssegmentering er også svært viktig for å vurdere behandlingsresponsen. Det er forskjellige metoder som blir brukt for bildesegmentering. De kan klassifiseres basert på hvordan de er formulert og hvordan segmenteringsprosessen utføres. Metodene inkluderer de som er baserte på terskelverdier, graf-baserte, kant-baserte, klynge-baserte, modell-baserte og hybride metoder, og metoder basert på maskinlæring og dyp læring. Andre metoder er baserte på å utvide, splitte og legge sammen regioner, å finne diskontinuiteter i randen, vannskille segmentering, aktive kontuter og graf-baserte metoder. I denne avhandlingen har vi utviklet metoder for å segmentere forskjellige typer medisinske bilder. Vi testet metodene på datasett for hvite blodceller (WBCs) og magnetiske resonansbilder (MRI). De utviklede metodene og analysen som er utført på bildedatasettet er presentert i tre artikler. I artikkel A (Paper A) foreslo vi en metode for segmentering av nukleuser og cytoplasma fra hvite blodceller. Metodene estimerer terskelen for segmentering av nukleuser automatisk basert på lokale minima. Metoden segmenterer WBC-ene før segmentering av cytoplasma avhengig av kompleksiteten til objektene i bildet. For bilder der WBC-ene er godt skilt fra røde blodlegemer (RBC), er WBC-ene segmentert ved å ta gjennomsnittet av $n$ bilder som allerede var filtrert med en terskelverdi. For bilder der RBC-er overlapper WBC-ene, er hele WBC-ene segmentert ved hjelp av enkle lineære iterative klynger (SLIC) og vannskillemetoder. Cytoplasmaet oppnås ved å trekke den segmenterte nukleusen fra den segmenterte WBC-en. Metoden testes på to forskjellige offentlig tilgjengelige datasett, og resultatene sammenlignes med toppmoderne metoder. I artikkel B (Paper B) foreslo vi en metode for segmentering av hjernesvulster basert på minste dekkende tre-konsepter (minimum spanning tree, MST). Metoden utfører interaktiv segmentering basert på MST. I denne artikkelen er bildet lastet inn i et interaktivt vindu for segmentering av svulsten. Fokusregion og bakgrunn skilles ved å klikke for å dele MST i to trær. Ett av disse trærne representerer fokusregionen og det andre representerer bakgrunnen. Den foreslåtte metoden ble testet ved å segmentere to forskjellige 2D-hjerne T1 vektede magnetisk resonans bildedatasett. Metoden er enkel å implementere og resultatene indikerer at den er nøyaktig og effektiv. I artikkel C (Paper C) foreslår vi en metode som behandler et 3D MRI-volum og deler det i hjernen, ikke-hjernevev og bakgrunnsegmenter. Det er en grafbasert metode som bruker MST til å skille 3D MRI inn i de tre regiontypene. Grafen lages av et forhåndsbehandlet 3D MRI-volum etterfulgt av konstrueringen av MST-en. Segmenteringsprosessen gir tre merkede, sammenkoblende komponenter som omformes tilbake til 3D MRI-form. Etikettene brukes til å segmentere hjernen, ikke-hjernevev og bakgrunn. Metoden ble testet på tre forskjellige offentlig tilgjengelige datasett og resultatene ble sammenlignet med ulike toppmoderne metoder.In image segmentation, an image is divided into separate objects or regions. It is an essential step in image processing to define areas of interest for further processing or analysis. The segmentation process reduces the complexity of an image to simplify the analysis of the attributes obtained after segmentation. It changes the representation of the information in the original image and presents the pixels in a way that is more meaningful and easier to understand. Image segmentation has various applications. For medical images, the segmentation process aims to extract the image data set to identify areas of the anatomy relevant to a particular study or diagnosis of the patient. For example, one can locate affected or abnormal parts of the body. Segmentation of follow-up data and baseline lesion segmentation is also very important to assess the treatment response. There are different methods used for image segmentation. They can be classified based on how they are formulated and how the segmentation process is performed. The methods include those based on threshold values, edge-based, cluster-based, model-based and hybrid methods, and methods based on machine learning and deep learning. Other methods are based on growing, splitting and merging regions, finding discontinuities in the edge, watershed segmentation, active contours and graph-based methods. In this thesis, we have developed methods for segmenting different types of medical images. We tested the methods on datasets for white blood cells (WBCs) and magnetic resonance images (MRI). The developed methods and the analysis performed on the image data set are presented in three articles. In Paper A we proposed a method for segmenting nuclei and cytoplasm from white blood cells. The method estimates the threshold for segmentation of nuclei automatically based on local minima. The method segments the WBCs before segmenting the cytoplasm depending on the complexity of the objects in the image. For images where the WBCs are well separated from red blood cells (RBCs), the WBCs are segmented by taking the average of $n$ images that were already filtered with a threshold value. For images where RBCs overlap the WBCs, the entire WBCs are segmented using simple linear iterative clustering (SLIC) and watershed methods. The cytoplasm is obtained by subtracting the segmented nucleus from the segmented WBC. The method is tested on two different publicly available datasets, and the results are compared with state of the art methods. In Paper B, we proposed a method for segmenting brain tumors based on minimum spanning tree (MST) concepts. The method performs interactive segmentation based on the MST. In this paper, the image is loaded in an interactive window for segmenting the tumor. The region of interest and the background are selected by clicking to split the MST into two trees. One of these trees represents the region of interest and the other represents the background. The proposed method was tested by segmenting two different 2D brain T1-weighted magnetic resonance image data sets. The method is simple to implement and the results indicate that it is accurate and efficient. In Paper C, we propose a method that processes a 3D MRI volume and partitions it into brain, non-brain tissues, and background segments. It is a graph-based method that uses MST to separate the 3D MRI into the brain, non-brain, and background regions. The graph is made from a preprocessed 3D MRI volume followed by constructing the MST. The segmentation process produces three labeled connected components which are reshaped back to the shape of the 3D MRI. The labels are used to segment the brain, non-brain tissues, and the background. The method was tested on three different publicly available data sets and the results were compared to different state of the art methods.Doktorgradsavhandlin

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

Micro/Nano Devices for Blood Analysis, Volume II

The development of micro- and nanodevices for blood analysis continues to be a growing interdisciplinary subject that demands the careful integration of different research fields. Following the success of the book “Micro/Nano Devices for Blood Analysis”, we invited more authors from the scientific community to participate in and submit their research for a second volume. Researchers from different areas and backgrounds cooperated actively and submitted high-quality research, focusing on the latest advances and challenges in micro- and nanodevices for diagnostics and blood analysis; micro- and nanofluidics; technologies for flow visualization and diagnosis; biochips, organ-on-a-chip and lab-on-a-chip devices; and their applications to research and industry

The Retinal Microvasculature in Secondary Progressive Multiple Sclerosis

In light of new data regarding pathology of multiple sclerosis (MS), more research is needed into the vascular aspects of the disease. Demyelination caused by inflammation is historically thought of as the main cause of disability in the disease. Recent studies, however, have suggested that MS is in fact a spectrum of overlapping phenotypes consisting of inflammation, oxidative damage and hypoperfusion. The microvasculature plays an important role in all of these pathogenic processes and its dysfunction may therefore be of crucial importance to the development and progression of the disease. This thesis focuses on investigating the microvasculature of the retina as a surrogate for the brain by assessing the vascular structure, blood flow dynamics and oxygen transfer of the retinal blood vessels in secondary progressive multiple sclerosis (SPMS). Studying the retinal microvasculature using a multimodal imaging approach has allowed us to develop a more detailed understanding of blood flow in MS and to identify new imaging markers for trials into neuroprotective drugs in MS. The work done in this thesis demonstrated; i) a higher rate of retinal microvascular abnormalities in MS which progresses with disease severity, ii) evidence of retinal vascular remodelling in SPMS and iii) changes in blood velocity and flow in the retina in SPMS. These observations pave the way for future investigations into the mechanisms of vascular alterations and vascular dysfunction in MS, and provide a set of imaging markers to further explore other cerebrovascular diseases through the retina