Automatic Counting Method Research Articles

A Hybrid Method for Automatic Counting of Microorganisms in Microscopic Images

Microscopic image analysis is an essential process to enable the automatic enumeration and quantitative analysis of microbial images. There are several system are available for numerating microbial growth. Some of the existing method may be inefficient to accurately count the overlapped microorganisms. Therefore, in this paper we proposed an efficient method for automatic segmentation and counting of microorganisms in microscopic images. This method uses a hybrid approach based on morphological operation, active contour model and counting by region labelling process. The colony count value obtained by this proposed method is compared with the manual count and the count value obtained from the existing method.

Evaluation of the interference of platelets in reticulocyte counting by flow cytometry using thiazole orange

Introduction: The reticulocyte count by flow cytometry (FC) - an automated counting method - can present errors due to the presence of interfering factors, contributing to a slight increase in results. However, automated methods have large advantages over the manual method, taken as reference, what justifies efforts to improve their quality. Objective: Evaluate platelet interference with the reticulocyte count by FC, using thiazole orange (TO) (FC/TO). Materials and methods: The method of reticulocyte count by FC/TO and a modified automated equivalent method, which excluded CD61-positive cells (platelets) from analysis (FC/TO/MOD), were compared to the manual method. Conclusion: Results were analyzed according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI) to assess interchangeability between the methods, by linear regression analysis and paired t-test. The exclusion of interfering fragments from result analysis by the modified method produced results in closer proximity to those of the reference method.

An automatic people counting method of hotel dining with occlusion

Video image has the advantage of large amount of information, good real-time performance and low cost, so automatic people counting based on video image has very high practical value, and many scholars have done a large number of experiments and studies on this and achieved certain achievements. But for scenes with more occlusion and background changing quickly and without obvious rules, it’s difficult to count accurately. In order to improve the counting accuracy in the above scenes, to provide the number of customers for hotel managers to efficiently organize and work, based on pictures, a automatic people counting method using SVM as weak classifiers, train intensively in learning by Adaboost algorithm(i.e. Adab_SVM algorithm) of hotel dining is proposed. The method is mainly aimed at the hotel scenes with occlusion too much to complete the segmentation of human body region. Firstly, traversing the entire picture to get the preliminary head areas and the number of people, then merge these head areas to get the exact number of people, to complete the statistical work on the number of people of the entire picture. Experimental results show that the method has higher counting accuracy in the hotel scenes with occlusion.

Automated image-processing for counting seedlings in a wheat field

Wheat field seedling density has a significant impact on the yield and quality of grains. Accurate and timely estimates of wheat field seedling density can guide cultivation to ensure high yield. The objective of this study was to develop an image-processing based, automatic counting method for wheat field seedlings, to investigate the principle of automatic counting of wheat emergence in the field, and to validate the newly developed method in various conditions. Digital images of the wheat fields at seedling stages with five cultivars and five seedling densities were acquired directly from above the fields. The wheat seedlings information was extracted from the background using excessive green and Otsu’s method. By analyzing the characteristic parameters of the overlapping regions (Overlapping region is a number of overlapping wheat seedlings in the image) of the fields, a chain code-based skeleton optimization method and corresponding equation were established for automatic counting of wheat seedlings in the overlapping regions. The results showed that the newly developed method can effectively count the number of wheat seedlings, with an average accuracy rate of 89.94 % and a highest accuracy rate of 99.21 %. The results also indicated that the accuracy of counting was not affected by different cultivars. However, the seedling density had significant impact on the counting accuracy (P 92 %) could be obtained. The study demonstrated that the newly developed method is reliable for automatic wheat seedlings counting, and also provides a theoretical perspective for automatic seedling counting in the wheat field.

Semi-automated identification of cones in the human retina using circle Hough transform.

A large number of human retinal diseases are characterized by a progressive loss of cones, the photoreceptors critical for visual acuity and color perception. Adaptive Optics (AO) imaging presents a potential method to study these cells in vivo. However, AO imaging in ophthalmology is a relatively new phenomenon and quantitative analysis of these images remains difficult and tedious using manual methods. This paper illustrates a novel semi-automated quantitative technique enabling registration of AO images to macular landmarks, cone counting and its radius quantification at specified distances from the foveal center. The new cone counting approach employs the circle Hough transform (cHT) and is compared to automated counting methods, as well as arbitrated manual cone identification. We explore the impact of varying the circle detection parameter on the validity of cHT cone counting and discuss the potential role of using this algorithm in detecting both cones and rods separately.

Counting White Blood Cells from a Blood Smear Using Fourier Ptychographic Microscopy.

White blood cell (WBC) count is a valuable metric for assisting with diagnosis or prognosis of various diseases such as coronary heart disease, type 2 diabetes, or infection. Counting WBCs can be done either manually or automatically. Automatic methods are capable of counting a large number of cells to give a statistically more accurate reading of the WBC count of a sample, but the specialized equipment tends to be expensive. Manual methods are inexpensive since they only involve a conventional light microscope setup. However, it is more laborious and error-prone because the small field-of-view (FOV) of the microscope necessitates mechanical scanning of a specimen for counting an adequate number of WBCs. Here, we investigate the use of Fourier ptychographic microscopy (FPM) to bypass these issues of the manual methods. With a 2x objective, FPM can provide a FOV of 120 mm2 with enhanced resolution comparable to that of a 20x objective, which is adequate for non-differentially counting WBCs in just one FOV. A specialist was able to count the WBCs in FPM images with 100% accuracy compared to the count as determined from conventional microscope images. An automatic counting algorithm was also developed to identify WBCs from FPM’s captured images with 95% accuracy, paving the way for a cost-effective WBC counting setup with the advantages of both the automatic and manual counting methods.

PWE-108 National survey of ascitic fluid wbc analysis method in the diagnosis of spontaneous bacterial peritonitis

Introduction Spontaneous bacterial peritonitis (SBP) commonly found in patients with cirrhosis and ascites has significant mortality (10%) hence the need for prompt diagnosis and appropriate therapy. Diagnosis of SBP is based on neutrophil count1in ascitic fluid >250/mm3, traditionally measured manually; however there is excellent correlation between manual and automated counting methods.2Anecdotally, UK hospitals use both techniques. This national survey aims to establish which methods are used currently to diagnose SBP, its diagnostic criteria and limitations. Method UK Hospitals (n = 164) were invited to participate in a survey to establish the method used to quantify ascitic fluid WBC in diagnosis of SBP: manual cell count performed by microbiologist or fully automated cell count. They were asked also to comment on specific cut-off points used for diagnosis of SBP, the speed of analysis and any other comments. Results Results from eight national UK liver transplant centres suggested a lack of consistency, with 50% using manual cell count and the other 50% using automation. A similar pattern was seen nationally (n = 100), with 66% using a manual technique and 34% an automated method. Timely WBC counts were often not obtained manually eg results “within 18 h”, ”overnight”, ”four hours to get result”. In some cases, samples were sent by taxi to another hospital for manual analysis. In contrast, ascitic fluid WBC counts were generally obtained within 30–60 min utilising automated analysis. Nationally, lack of consensus was apparent in the cut-off used to diagnose SBP, with many hospitals using total WBC >250/mm3, rather than neutrophil count >250/mm3. In some hospitals, entirely different cut-offs were used, including: total WBC >500/mm3; 250 WBC/mm3or 150/mm3neutrophils; WBC >300/mm3; WBC >400/mm3or >250/mm3neutrophils. Some hospitals (n = 4) used additionally urine dipstick analysis of ascitic fluid. The cost of manual WBC ascitic fluid count is estimated at £16.83, whilst the cost of an automated count is £3.53, suggesting that nationally considerable cost savings could be made if all ascitic fluid samples are analysed in an automated way. Conclusion This national survey has highlighted the lack of consistency in the measurement of ascitic fluid WBC in the diagnosis of SBP across the UK, both in terms of method utilised and WBC cut-offs. Automated analysis produced faster results (<1 hr) and allowed differential WBC (incl neutrophil count) to be obtained readily. Widespread uptake of automated ascitic WBC measurement in conjunction with a clear WBC cut-off, as exemplified by use of the decompensated cirrhosis care bundle3is likely to improve patient care and outcomes. Disclosure of interest None Declared.

Automatic enumeration of gold nanomaterials at the single-particle level.

In this study, we developed a highly sensitive automatic counting method for gold nanomaterials at the single particle level, which can serve as a general sensing platform based on counting of gold nanomaterials. This method substantially improved the sensitivity and accuracy for AuNP counting by adopting the color image processing based on the distinctive localized plasmonic light-scattering of gold nanomaterials. The 60-nm AuNPs, with concentrations down to 4 fM, can be detected with our method. As a universal counting approach for gold nanomaterials, such as gold nanospheres, nanorods, and aggregates from particles under detectable size, this quantification method should be versatile to a breadth of applications.

Partial Least Squares回帰及び複数特徴量間HLAC特徴を用いた細胞内画像中の粒子計数

In the field of cell biology, particle counting in intracellular images is important for investigating the cause of diseases. However, particles are manually counted by human observers now. Such manually counting takes a lot of time, and counting result becomes subjective. If an automatic counting method by computer is realized, we can treat a large number of images and it can obtain many objective data. The development of an automatic counting method much contributes to understand the case of disease. However, particle counting in intracellular images by a computer is new research field, and conventional methods are little. Thus, we propose counting method based on regression analysis. We use partial least squares regression and auto-correlation between 2 different types of features using mask patterns for higher-order local autocorrelation feature. The proposed method gives higher accuracy than counting by principal component regression, support vector regression and ImageJ. In experiment, the proposed method can count with small error in comparison with human counting. The effectiveness of our method is shown by experiments.

On XML Based Automated Function Point Analysis: An Effective Method to Assess Developer Productivity

This article proposes a novel approach to objectively measure developer productivity and individual contribution to development projects using automated function point analysis (integrated with source control system (SCS) and continuous integration (CI)). Automated function point analysis is a central tenant required in this approach. The method proposed herein relies on pattern matching to identify and group fields in to internal logical files, external inputs, external outputs, and external query objects which can be used to identify the objective size of software. Unlike earlier automated functional sizing algorithms, the method proposed is applicable to any application source code that expresses its user interface in well-formed extensible markup language (XML). This study shows that a prototype implementation of the proposed automated counting method is capable of providing a reliable and consistent relative size of existing software (accurate to +/-6.65% and 5000 times faster), and goes on to explain how this method can be integrated with existing requirements engineering tools, source control systems and continuous integration tooling in order to produce an objective measure of software developer output compared to estimates established during requirements gathering and planning that exceeds the common, subjective means that are in use in many organizations today.

Particle detection in intracellular images and radius estimation by circle fitting

Detecting particles in intracellular images and estimating their radii is important for investigating the causes of clinical conditions. Presently, particles are manually counted by observers, which is time consuming and subjective. Thus, we propose an automatic particle counting and radius estimation method based on pattern recognition techniques. Particles are analyzed by a computer in two processes. During the first process, particles are detected using face recognition via a support vector machine. In the second process, the edge points of a detected particle are distinguished by the intensity difference between the particle and the background. The edge points are extracted and fitted by a circle by a voting technique. The proposed method accurately located 96.04% of the particles detected by manual counting. ImageJ, the particle detection software generally used in cell biology, detected 72.32% of the particles. Therefore, our method yields higher accuracy than ImageJ. Moreover, the effectiveness of radius estimation was also experimentally verified. © 2014 Institute of Electrical Engineers of Japan. Published by John Wiley &amp; Sons, Inc.

An Automatic Microalgal Cells Counting Method

An automatic counting method for microalgal cells were proposed using digital image processing with characteristics of microalgae microscopic image considered. Firstly, the microalgae image was pretreated with graying, and was then applied the median filtering to remove the noise. Secondly, the image was applied Bot-hat conversion to enhance the contrast. Thirdly, the image was segmented with Otsu algorithm, and was then applied morphological operations. Finally, the binary image segments representing microalgal cells were labeled and counted. The results of experiments showed that this method was simple, efficient and accurate in counting microalgal cells in the microscopic image.

Middle Jurassic coccolith fluxes: A novel approach by automated quantification

Coccolithophores, major pelagic carbonate producers, underwent important changes during the latest Aalenian–Early Bajocian (ca. − 170 Ma, Middle Jurassic). Specifically, a new genus called Watznaueria diversified and started dominating the coccolith assemblages. While this change in the coccolith assemblages is already known, the impact on coccolith absolute abundances and fluxes is still misunderstood. Here we tested for the first time on Mesozoic deposits an automated counting method using the SYRACO system, and then compared these results with a classic counting method. Two neural networks were created based on the shape, birefringence and size of coccoliths, the first of which contained only one class and the second of which was composed of six classes. Based on the morphological criteria used for the neural networks, coccoliths were not identified at the species level. We quantified coccolith absolute abundances and estimated fluxes for the latest Aalenian–Early Bajocian in two well-dated and correlated sections, Cabo Mondego in Portugal and Chaudon-Norante in France. There is a strong linear correlation between classic and automatic counting (r > 0.8) supporting the latter as a reliable method for coccolith absolute abundance and flux quantification. In spite of under- or over-estimation by automatic counting with respect to classic counting, the absolute abundance stratigraphic trends are remarkably similar. Under- or over-estimation may be reduced by further technical improvements such as automatic focus. At Cabo Mondego, coccolith fluxes increased from 0.2 × 109 coccoliths/m2/yr in the latest Aalenian to 700 × 109 coccoliths/m2/yr in the middle of the Laeviuscula ammonite Zone (Early Bajocian), remaining high for the rest of the Early Bajocian. At Chaudon-Norante, a similar trend was observed. This coccolith flux increase is therefore considered to be a supra-regional event matching an important δ13CBulk carbonate positive excursion. The carbon isotopic positive excursion, along with the onset of radiolarian sedimentation and coccolith fluxes is related to a gradual increase in the fertility of the oceans during the Early Bajocian and is likely associated with warmer and more humid climates and associated enhanced continental weathering.

Automated Versus Manual Platelet Count in Aden

This is a cross sectional study conducted in the National Center of Public Health Laboratories (NCPHL) of Aden – Yemen, during October-December 2011, with an overall aim to compare the results of platelet count by the automated versus the manual methods. Study included 190 random samples, examined with confirmatory platelet estimate done by the stained thin blood film. The mean platelets count estimated by the manual method was 225.2 ± 95.4 × 109/L, while that estimated by the automated method was 245.7 ± 109.8 × 109/L, with no significant statistical difference between both means (p>0.05). The Pearson correlation test showed significant positive correlation between both methods (r: 0.563), this correlation remained significant when the samples of normal count by the two methods were correlated (r: 0.359), but it was insignificant negative correlation when the samples of low or high counts by the two methods were correlated (r: -0.151 and -0.559) respectively. This study concluded that significant positive correlation is present between the manual and the automated counting methods of platelets and recommended that platelet count is not varied when done by manual or automated methods, but in every method, it should be accompanied by platelet estimate, especially with abnormal counts.

Mir143 expression inversely correlates with nuclear ERK5 immunoreactivity in clinical prostate cancer

Background:Aberrant mitogen/extracellular signal-regulated kinase 5 (MEK5)–extracellular signal-regulated protein kinase 5 (ERK5)-mediated signalling has been implicated in a number of tumour types including prostate cancer (CaP). The mechanism for ERK5 activation in CaP remains to be fully elucidated. Studies have recently implicated the role of microRNA (miRNA) mir143 expression in the regulation of ERK5 expression.Methods:We utilised a tissue microarray (TMA) of 530 CaP cores from 168 individual patients and stained for both mir143 and ERK5. These TMAs were scored by a combination of observer and automated methods.Results:We observed a strong inverse relation between ERK5 and mir143, which manifested itself most strongly in the subgroup of 417 cores with non-zero mir143 and ERK5 immunoreactivity, or with only one of mir143 or ERK5 being zero (cc=0.2558 and P<0.0001). Mir143 neither correlate with Gleason scores or prostate-specific antigen levels, nor was it a predictor of disease-specific survival on univariate analysis.Conclusion:Although the mechanism for ERK5 activation in CaP remains to be fully elucidated, we have further validated the potential role of mir143 in regulating ERK5 levels in the clinical context. In addition, we demonstrate that the automated counting method for nuclear ERK5 is a clinically useful alterative to observer counting method in patient stratification in the context of ERK5 targeting therapy.

An improved respiratory syncytial virus neutralization assay based on the detection of green fluorescent protein expression and automated plaque counting

BackgroundVirus neutralizing antibodies against respiratory syncytial virus (RSV) are considered important correlates of protection for vaccine evaluation. The established plaque reduction assay is time consuming, labor intensive and highly variable.MethodsHere, a neutralization assay based on a modified RSV strain expressing the green fluorescent protein in combination with automated detection and quantification of plaques is described.ResultsThe fluorescence plaque reduction assay in microplate format requires only two days to complete and is simple and reproducible. A good correlation between visual and automated counting methods to determine RSV neutralizing serum antibody titers was observed.ConclusionsThe developed virus neutralization assay is suitable for high-throughput testing and can be used for both animal studies and (large scale) vaccine clinical trials.

Automatic neutrophil nucleus lobe counting based on graph representation of region skeleton

Abnormal neutrophil nucleus lobation (like left shift and right shift) helps to diagnose for some clinical conditions. Currently, quantification of it depends on the manual microscopic inspection of blood smears by clinicians. The quality of the manual inspection is extremely limited by the efficiency of clinicians and their medical background. This article proposed an automatic lobe counting method based on the graph representation of the nucleus region skeletons. Skeletons of the segmented nucleus regions are computed by augmented Fast Marching Method and transformed into plane graphs. Then the nucleus lobes are split based on the extracted graph properties including width distribution along the skeleton and graph structure decomposition. Experiments show that the proposed method could efficiently approaches the real lobe numbers in blood smears and reliably distinguish the stabs from the segmented neutrophils, thus it should be helpful in automatic neutrophil lobe number quantification and abnormal lobation diagnosis.

Counting passing people with different ways of movement in horizontal view

AbstractCounting of passing people in a passage or corridor is considered important for security and in various kinds of investigations. This paper proposes a method for automatic counting of moving people with the different ways of movement in a horizontal view. The centers of gravity of motion detection blocks are plotted in the plotting space, and the movement of people can be detected by the Hough transform without motion tracking. The intercept and the slope of a detected line represent the starting time when a person enters the scene and the speed of movement of the target. The ways of movement also can be recognized by using the difference in the speeds of movement of detected persons. Experimental results showed that reliable detection and counting of passing people were realized by the proposed method. © 2011 Wiley Periodicals, Inc. Electron Comm Jpn, 94(9): 1–9, 2011; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.10363

Chronic and Acute Models of Retinal Neurodegeneration TrkA Activity Are Neuroprotective whereas p75NTR Activity Is Neurotoxic through a Paracrine Mechanism

In normal adult retinas, NGF receptor TrkA is expressed in retinal ganglion cells (RGC), whereas glia express p75(NTR). During retinal injury, endogenous NGF, TrkA, and p75(NTR) are up-regulated. Paradoxically, neither endogenous NGF nor exogenous administration of wild type NGF can protect degenerating RGCs, even when administered at high frequency. Here we elucidate the relative contribution of NGF and each of its receptors to RGC degeneration in vivo. During retinal degeneration due to glaucoma or optic nerve transection, treatment with a mutant NGF that only activates TrkA, or with a biological response modifier that prevents endogenous NGF and pro-NGF from binding to p75(NTR) affords significant neuroprotection. Treatment of normal eyes with an NGF mutant-selective p75(NTR) agonist causes progressive RGC death, and in injured eyes it accelerates RGC death. The mechanism of p75(NTR) action during retinal degeneration due to glaucoma is paracrine, by increasing production of neurotoxic proteins TNF-α and α(2)-macroglobulin. Antagonists of p75(NTR) inhibit TNF-α and α(2)-macroglobulin up-regulation during disease, and afford neuroprotection. These data reveal a balance of neuroprotective and neurotoxic mechanisms in normal and diseased retinas, and validate each neurotrophin receptor as a pharmacological target for neuroprotection.

Comparison of endothelial cell loss after cataract surgery: Phacoemulsification versus manual small-incision cataract surgery: Six-week results of a randomized control trial

To compare endothelial cell loss in cataract surgery by phacoemulsification and by manual small-incision cataract surgery (SICS). Tertiary care ophthalmic center, India. A complete ophthalmologic examination and endothelial specular microscopy were performed preoperatively and 1 and 6 weeks postoperatively in patients having cataract surgery. The endothelial cell count (ECC) was calculated manually and automatically using an LSM 12000 specular microscope. Patients were randomly allocated to have SICS or phacoemulsification using a random number table. Phacoemulsification was performed using the stop-and-chop technique and SICS, by viscoexpression. The study evaluated 200 patients, 100 in each group. The mean preoperative ECC by the manual counting method was 2950.7 cells/mm(2) in the phacoemulsification group and 2852.5 cells/mm(2) in the SICS group and by the automated counting method, 3053.7 cells/mm(2) and 2975.3 cells/mm(2), respectively. The difference at 6 weeks was 543.4 cells/mm(2) and 505.9 cells/mm(2), respectively, by the manual method (P = .44) and 474.2 cells/mm(2) and 456.1 cells/mm(2), respectively, by the automated method (P = .98). The corrected distance visual acuity at 6 weeks was better than 6/18 in 98.5% of eyes in the phacoemulsification group and 97.3% of eyes in the SICS group. There were no clinically or statistically significant differences in ECC loss or visual acuity between phacoemulsification and SICS, although there was a small difference in the astigmatic shift.