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[ACM Press the 3rd International Conference - Seoul, Republic of Korea (2018.08.22-2018.08.24)] Proceedings of the 3rd International Conference on Biomedical Signal and Image Processing - ICBIP '18 - Automatic Detection of Cell Regions in Microscope Images Based on BFED Algorithm
摘要： Circulating tumor cells (CTC) attract attention as a biomarker that can evaluate cancer metastasis and therapeutic effects. The CTC exists in the blood of cancer patients, so pathologists analyze blood by using a fluorescence microscope. However, manual analysis by pathologists is hard-work since the number of CTC to substances contained in the blood is very few and the cell regions are often unclear depending on shooting environments. In addition, there are few studies on automatic identification of CTC. In this paper, we develop an automatic detection method of cell regions in microscope images based on bacterial foraging-based edge detection (BFED) algorithm to analyze CTC. In the first step, we detect the initial cell regions by BFED algorithm. Second, we identify whether the region is a single cell or multiple cells come in connect with other cell(s) by SVM. Third, when a cell is connected with other one, we separate the connecting cells by branch and bound algorithm and obtain the final cell regions. We applied our proposed method to 1680 microscopy images (6 cases). The experimental results demonstrate that the proposed method has a true positive rate of 93.9% and a false positive 1.29 /case.
关键词： Saliency map,Computer aided diagnosis,Support vector machine,Branch and bound algorithm,Circulating tumor cells,BFED algorithm
更新于2025-09-23 15:22:29
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Detection of Circulating Tumor Cells in Fluorescence Microscopy Images Based on ANN Classifier
摘要： Circulating tumor cells (CTCs) is a clinical biomarker for cancer metastasis. CTCs are cells circulating in the body of patients by being separated from primary cancer and entering into blood vessel. CTCs spread every positions in the body, and this is one of the cause of cancer metastasis. To analyze them, pathologists get information about metastasis without invasive test. CTCs test is conducted by analyzing the blood sample from patient. The fluorescence microscope generates a large number of images per each sample, and images contain a lot of cells. There are only a few CTCs in images and cells often have blurry boundaries. So CTCs identification is not an easy work for pathologists. In this paper, we develop an automatic CTCs identification method in fluorescence microscopy images. This proposed method has three section. In the first approach, we conduct the cell segmentation in images by using filtering methods. Next, we compute feature values from each CTC candidate region. Finally, we identify CTCs using artificial neural network algorithm. We apply the proposed method to 5895 microscopy images (7 samplesas), and evaluate the effectiveness of our proposed method by using leave-one-out cross validation. We achieve the result of performance tests, a true positive rate is 92.57% and false positive rate is 9.156%.
关键词： Fluorescence microscopy image,Artificial neural network,Feature analysis,Computer aided diagnosis,Circulating tumor cells
更新于2025-09-23 15:22:29
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Sensing biomarkers with plasmonics
摘要： The detection of biomarkers is critical for enabling early disease diagnosis, monitoring the progression, and tracking the effectiveness of therapeutic intervention. Plasmonic sensors exhibit a broad range of analytical capabilities, from the rapid generation of colorimetric readouts to single-molecule sensitivity in ultralow sample volumes, which have led to their increased exploration in bioanalysis and point-of-care applications. This perspective presents selected accounts of recent developments on the different types of plasmonic sensing platforms, the pervasive challenges and outlook on the pathway to translation. We highlight the sensing of upcoming biomarkers including microRNA, circulating tumor cells, exosomes, and cell-free DNA, and discuss the opportunity of utilizing plasmonic nanomaterials and tools for biomarker detection beyond biofluids, such as in tissues, organs, and disease sites. The integration of plasmonic biosensors with established and upcoming technologies of instrumentation, sample pre-treatment, and data analysis will help realize their translation to clinical settings for improving healthcare and enhancing the quality of life.
关键词： plasmonics,exosomes,biomarkers,microRNA,biosensors,circulating tumor cells,cell-free DNA
更新于2025-09-23 15:21:01
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Live circulating tumour cells selection on digitized self-assembled cell array (Digi-saca) chip by in-parallel/in-situ image analysis, cell capture, and cultivation
摘要： Cancer is one of the major and most deadly diseases of mankind. With present technologies, early detection and prevention of cancer disease is still a major bottleneck. Circulating Tumor Cells (CTC) is one of the cancer biomarkers for the diagnosis of metastasis. However, there is an urgent need to improve the turn around time of CTC detection from a small sample volume in order to promote its clinical implications. This paper highlights Digi-SACA, an automated workflow to capture CTCs from the whole blood on self-assembled cell array chips. The gravity force and lateral driving force based microfluidic chip drive the mononuclear blood cells from 4 ml of samples to form a monolayer without any external fluid control equipment. The subsequent immunostaining and automated image acquisition, image processing, CTC enumeration, and CTC harvest can be completed within 4 hours. The sensitivity of Digi-SACA chip is 1 in ten million leukocytes which is very promising for early detection of the CTC. In a small pilot study series, the Digi-SACA chip is able to detect of the mean of 14.4 CTCs from 4ml of blood in 10 clinical samples from patients with breast cancer, while mean of 17.6 CTCs were measured from the same sample set with IsoFluXTM platform from 4ml of blood. We have also demonstrated the efficiency of our technique to pick CTC with glass micropipettes for standard cell culture growth up to 11-15 days. Overall, our study represents the liability of the Digi-SACA system in CTCs enumeration, detection, and isolation against the state-of-the-art.
关键词： Breast cancer,Digi-SACA chip,Single-cell picking,Circulating tumor cells (CTC)
更新于2025-09-23 15:19:57
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Bioinspired magnetic nanoparticles as multimodal photoacoustic, photothermal and photomechanical contrast agents
摘要： Nanoparticles from magnetotactic bacteria have been used in conventional imaging, drug delivery, and magnetic manipulations. Here, we show that these natural nanoparticles and their bioinspired hybrids with near-infrared gold nanorods and folic acid can serve as molecular high-contrast photoacoustic probes for single-cell diagnostics and as photothermal agents for single-cell therapy using laser-induced vapor nanobubbles and magnetic field as significant signal and therapy amplifiers. These theranostics agents enable the detection and photomechanical killing of triple negative breast cancer cells that are resistant to conventional chemotherapy, with just one or a few low-energy laser pulses. In studies in vivo, we discovered that circulating tumor cells labeled with the nanohybrids generate transient ultrasharp photoacoustic resonances directly in the bloodstream as the basis for new super-resolution photoacoustic flow cytometry in vivo. These properties make natural and bioinspired magnetic nanoparticles promising biocompatible, multimodal, high-contrast, and clinically relevant cellular probes for many in vitro and in vivo biomedical applications.
关键词： theranostics,super-resolution,contrast agents,photomechanical,photoacoustic,magnetic nanoparticles,photothermal,circulating tumor cells,bioinspired,magnetotactic bacteria
更新于2025-09-19 17:15:36
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Multifunctional luminescent immuno-magnetic nanoparticles: toward fast, efficient, cell-friendly capture and recovery of circulating tumor cells
摘要： Highly efficient isolation and recovery of viable circulating tumor cells (CTCs) from the blood of patients is an important precondition to address the current dilemma of insufficient CTC studies, and can promote the development of individualized antitumor therapies. Herein, a cell-friendly CTC isolation and recovery nanoplatform with luminescent labelling was established using a layer-by-layer (LbL) assembly technique and a stimulated cellular-release strategy. In particular, the anti-epithelial cell adhesion molecule (anti-EpCAM) antibody was introduced with a disulfide bond-containing linker for further bio-friendly recovery of the CTCs. Quantum dots (QDs) were deposited onto fast magnet-responsive Fe3O4 nanoparticles through a facile LbL assembly method to monitor the capture and recovery process in real time. The obtained PEGylated immuno-magnetic nanospheres (PIMNs) can all be magnetically collected within 2 min. Capture efficiencies above 90% can be achieved from blood samples with 5–200 CTCs per mL after only 1–2 min incubation. Nearly all PIMNs on the surface of the CTCs were detached after 15 min of glutathione (GSH) treatment with the disappearance of QD signals. Recovered CTCs could be directly used for culture (cell viability, 98%), and their invasiveness and migration characteristics remained unchanged. Furthermore, the PIMNs were successfully applied to isolate CTCs in cancer patients’ peripheral blood samples, and an average of 8.6 ± 5.8 CTCs per mL was detected. The results above suggested that PIMNs may serve as a powerful nanoplatform for CTC screening, isolation and recovery.
关键词： layer-by-layer assembly,circulating tumor cells,capture and recovery,Multifunctional luminescent immuno-magnetic nanoparticles,quantum dots,glutathione treatment
更新于2025-09-19 17:15:36
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Optofluidic real-time cell sorter for longitudinal CTC studies in mouse models of cancer
摘要： Circulating tumor cells (CTCs) play a fundamental role in cancer progression. However, limited blood volume and the rarity of CTCs in the bloodstream preclude longitudinal, in-depth studies of these cells using existing liquid biopsy techniques. Here, we present an optofluidic system that continuously collects fluorescently labeled CTCs from a genetically engineered mouse model (GEMM) for several hours per day over multiple days or weeks. The system is based on a microfluidic cell sorting chip connected serially to an unanesthetized mouse via an implanted arteriovenous shunt. Pneumatically controlled microfluidic valves capture CTCs as they flow through the device, and CTC-depleted blood is returned back to the mouse via the shunt. To demonstrate the utility of our system, we profile CTCs isolated longitudinally from animals over 4 days of treatment with the BET inhibitor JQ1 using single-cell RNA sequencing (scRNA-Seq) and show that our approach eliminates potential biases driven by intermouse heterogeneity that can occur when CTCs are collected across different mice. The CTC isolation and sorting technology presented here provides a research tool to help reveal details of how CTCs evolve over time, allowing studies to credential changes in CTCs as biomarkers of drug response and facilitating future studies to understand the role of CTCs in metastasis.
关键词： single-cell RNA-Seq,metastasis,circulating tumor cells,GEMM,microfluidic
更新于2025-09-19 17:15:36
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[IEEE 2019 Conference on Lasers and Electro-Optics Europe & European Quantum Electronics Conference (CLEO/Europe-EQEC) - Munich, Germany (2019.6.23-2019.6.27)] 2019 Conference on Lasers and Electro-Optics Europe & European Quantum Electronics Conference (CLEO/Europe-EQEC) - In Flow Manipulation and Characterization of Cancer Cells by Coherent Computational Microscopy
摘要： Liquid biopsy has shown remarkably promising in oncology for the early diagnosis of cancer through the detection of circulating biomarkers such as circulating tumor cells (CTCs). Recent evidences suggest that CTCs represent effective prognostic and predictive biomarkers to monitor/predict therapy efficacy in breast, colon and prostate cancers [1,2]. However, the frequency of CTCs in blood is approximately 1 to 10 cells per 10 mL of blood, which is as challenging as looking for a needle in a haystack. In microfluidics, Digital Holography (DH) has been shown to be a promising technique to characterize CTCs with the aim to detect them inside a heterogeneous liquid sample. DH is label-free, real-time and gives access to the complex amplitude of the object [3-6]. Thus, any classification approach based on the holographic signature can exploit a reach information content to take a decision. Moreover, the flexible refocusing capability of DH imaging allows to inspect an entire liquid volume with a single capture. This enables the high-throughput inspection of blood and other bodily fluids rapidly flowing inside microfluidic channels. In DH, the sample is probed from one single direction and the phase delay introduced by the sample in through transmission acts as a contrast agent. Hence, the optical thickness measurable by DH imaging is an integral information, i.e. the sum of all the contributions experienced by the coherent light during its passage through the sample. In order to decouple the refractive index from the physical thickness and to resolve its distribution along the optical axis, tomography exploits multiple recordings, probing the sample from different angles and combining the corresponding phase-contrast maps [7,8]. Various schemes have been proposed to minimize the number of sampling angles and to make the recording stage faster in order to match the requirements and time constraints imposed by real biological problems. Here we show the recent advances of in-flow holographic tomography, which exploits a controlled induced rotation of the sample inside the microfluidic channel to probe it from different view angles with no mechanical rotation of the source beam [8,9]. We introduce an effective algorithm to recover from the recorded phase maps the set of angles required as input of the optical projection tomography algorithm [7-9]. We show the application of holographic flow tomography to the characterization of different cancer cells [10], namely breast cancer cells, ovarian cancer cells and neuroblastoma. We also discuss different possibilities of Lab-on-a-Chip design and flow engineering that allow us to induce controlled rotations while maintaining the high-throughput nature of DH microscopy [9,11,12]. In the next future, the large amount of data obtainable by this approach will be used to train a neural network devoted to classify CTCs, distinguishing them from the other components of a blood stream.
关键词： liquid biopsy,cancer cells,microfluidics,circulating tumor cells,holographic tomography,Lab-on-a-Chip,digital holography
更新于2025-09-16 10:30:52
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Detection of Apoptotic Circulating Tumor Cells Using in vivo Fluorescence Flow Cytometry
摘要： Most cancer patients die from metastatic disease as a result of a circulating tumor cell (CTC) spreading from a primary tumor through the blood circulation to distant organs. Many studies have demonstrated the tremendous potential of using CTC counts as prognostic markers of metastatic development and therapeutic efficacy. However, it is only the viable CTCs capable of surviving in the blood circulation that can create distant metastasis. To date, little progress has been made in understanding what proportion of CTCs is viable and what proportion is in an apoptotic state. Here, we introduce a novel approach toward in situ characterization of CTC apoptosis status using a multi-color in vivo flow cytometry platform with fluorescent detection for the real-time identification and enumeration of such cells directly in blood flow. The proof of concept was demonstrated with two-color fluorescence flow cytometry (FFC) using breast cancer cells MDA-MB-231 expressing green fluorescein protein (GFP), staurosporine as an activator of apoptosis, Annexin-V apoptotic kit with orange dye color, and a mouse model. The future application of this new platform for real-time monitoring of antitumor drug efficiency is discussed.
关键词： fluorescence flow cytometry,circulating tumor cells,in vivo detection of circulating tumor cells,apoptosis
更新于2025-09-10 09:29:36
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DEPArray? system: An automatic image‐based sorter for isolation of pure circulating tumor cells
摘要： Circulating tumor cells (CTCs) are rare cells shed into the bloodstream by invasive tumors and their analysis offers a promising noninvasive tool to predict and monitor therapeutic responses. CTCs can be isolated from patient blood and their characterization at single-cell level can inform on the genomic landscape of a tumor. All CTC enrichment methods bear a burden of contaminating normal cells, which mandate a further step of purification to enable reliable downstream genetic analysis. Here, we describe the DEPArray? technology, a microchip-based digital sorter, which combines precise microfluidic and microelectronic enabling precise, image-based isolation of single CTCs, which can then be analyzed by Next Generation Sequencing (NGS) methods.
关键词： rare cells,pure cells,cancer cells,liquid biopsy,single cells,circulating tumor cells,image-based sorting
更新于2025-09-04 15:30:14