Dataset Information

DEPArray™ system: An automatic image-based sorter for isolation of pure circulating tumor cells.

ABSTRACT: 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. © 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.

SUBMITTER: Di Trapani M

PROVIDER: S-EPMC6590341 | biostudies-literature | 2018 Dec

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

DEPArray™ system: An automatic image-based sorter for isolation of pure circulating tumor cells.

Di Trapani Mariano M Manaresi Nicolò N Medoro Gianni G

Cytometry. Part A : the journal of the International Society for Analytical Cytology 20181201 12

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, w ...[more]

PMID: 30551261

Similar Datasets

Project description:Isolation of circulating tumor cells (CTCs) from peripheral blood has the potential to provide a far easier "liquid biopsy" than tumor tissue biopsies, to monitor tumor cell populations during disease progression and in response to therapies. Many CTC isolation technologies have been developed. We optimized the Parsortix system, an epitope independent, size and compressibility-based platform for CTCs isolation, making it possible to harvest CTCs at the speed and sample volume comparable to standard CellSearch system. We captured more than half of cancer cells from different cancer cell lines spiked in blood samples from healthy donors using this system. Cell loss during immunostaining of cells transferred and fixed on the slides is a major problem for analyzing rare cell samples. We developed a novel cell transfer and fixation method to retain >90% of cells on the slide after the immunofluorescence process without affecting signal strength and specificity. Using this optimized method, we evaluated the Parsortix system for CTC harvest in prostate cancer patients in comparison to immunobead based CTC isolation systems IsoFlux and CellSearch. We harvested a similar number (p = 0.33) of cytokeratin (CK) positive CTCs using Parsortix and IsoFlux from 7.5 mL blood samples of 10 prostate cancer patients (an average of 33.8 and 37.6 respectively). The purity of the CTCs harvested by Parsortix at 3.1% was significantly higher than IsoFlux at 1.0% (p = 0.02). Parsortix harvested significantly more CK positive CTCs than CellSearch (p = 0.04) in seven prostate cancer patient samples, where both systems were utilized (an average of 32.1 and 10.1 respectively). We also captured CTC clusters using Parsortix. Using four-color immunofluorescence we found that 85.8% of PC3 cells expressed EpCAM, 91.7% expressed CK and 2.5% cells lacked both epithelial markers. Interestingly, 95.6% of PC3 cells expressed Vimentin, including those cells that lacked both epithelial marker expression, indicating epithelial-to-mesenchymal transition. CK-positive/Vimentin-positive/CD45-negative, and CK-negative/Vimentin-positive/CD45-negative cells were also observed in four of five prostate cancer patients but rarely in three healthy controls, indicating that Parsortix harvests CTCs with both epithelial and mesenchymal features. We also demonstrated using PC3 and DU145 spiking experiment that Parsortix harvested cells were viable for cell culture.

Project description:Mosquito-borne diseases such as dengue fever and malaria are the top 10 leading causes of death in low-income countries. Control measure for the mosquito population plays an essential role in the fight against the disease. Currently, several intervention strategies; chemical-, biological-, mechanical- and environmental methods remain under development and need further improvement in their effectiveness. Although, a conventional entomological surveillance, required a microscope and taxonomic key for identification by professionals, is a key strategy to evaluate the population growth of these mosquitoes, these techniques are tedious, time-consuming, labor-intensive, and reliant on skillful and well-trained personnel. Here, we proposed an automatic screening, namely the deep metric learning approach and its inference under the image-retrieval process with Euclidean distance-based similarity. We aimed to develop the optimized model to find suitable miners and suggested the robustness of the proposed model by evaluating it with unseen data under a 20-returned image system. During the model development, well-trained ResNet34 are outstanding and no performance difference when comparing five data miners that showed up to 98% in its precision even after testing the model with both image sources: stereomicroscope and mobile phone cameras. The robustness of the proposed-trained model was tested with secondary unseen data which showed different environmental factors such as lighting, image scales, background colors and zoom levels. Nevertheless, our proposed neural network still has great performance with greater than 95% for sensitivity and precision, respectively. Also, the area under the ROC curve given the learning system seems to be practical and empirical with its value greater than 0.960. The results of the study may be used by public health authorities to locate mosquito vectors nearby. If used in the field, our research tool in particular is believed to accurately represent a real-world scenario.

Dataset Information

DEPArray™ system: An automatic image-based sorter for isolation of pure circulating tumor cells.

Publications

DEPArray™ system: An automatic image-based sorter for isolation of pure circulating tumor cells.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets