High-Throughput Automated Microscopy of Circulating Tumor Cells

Aguilar-Avelar, Carlos; Soto-García, Brenda; Araíz-Hernández, Diana; León, J.; Esparza, Miguel; Chacón, Franco; Delgado-Balderas, Jesús Rolando; Álvarez, Mario Moisés; Santiago, Grissel Trujillo‐de; Gómez‐Guerra, Lauro Salvador; Velarde-Calvillo, Liza P.; Abarca-Blanco, Alejandro; Wong-Campos, J. D.

doi:10.1038/s41598-019-50241-w

Cited by 16 publications

(13 citation statements)

References 37 publications

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“…In this study, cells categorized as CTCs were automatically classified by the algorithm implemented in the imaging system, these events were further corroborated by a trained operator. A detailed description and validation of the imaging system performance can be found in Aguilar-Avelar et al 45 . In addition, if molecular analysis are required, the membrane can be collected in a microtube for nucleic acid extraction using commercially available kits.…”

Section: Resultsmentioning

confidence: 99%

Characterization of a novel automated microfiltration device for the efficient isolation and analysis of circulating tumor cells from clinical blood samples

León¹,

Soto-García²,

Araíz-Hernández³

et al. 2020

Sci Rep

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The detection and analysis of circulating tumor cells (CTCs) may enable a broad range of cancer-related applications, including the identification of acquired drug resistance during treatments. However, the non-scalable fabrication, prolonged sample processing times, and the lack of automation, associated with most of the technologies developed to isolate these rare cells, have impeded their transition into the clinical practice. This work describes a novel membrane-based microfiltration device comprised of a fully automated sample processing unit and a machine-vision-enabled imaging system that allows the efficient isolation and rapid analysis of CTCs from blood. The device performance was characterized using four prostate cancer cell lines, including PC-3, VCaP, DU-145, and LNCaP, obtaining high assay reproducibility and capture efficiencies greater than 93% after processing 7.5 mL blood samples spiked with 100 cancer cells. Cancer cells remained viable after filtration due to the minimal shear stress exerted over cells during the procedure, while the identification of cancer cells by immunostaining was not affected by the number of non-specific events captured on the membrane. We were also able to identify the androgen receptor (AR) point mutation T878A from 7.5 mL blood samples spiked with 50 LNCaP cells using RT-PCR and Sanger sequencing. Finally, CTCs were detected in 8 out of 8 samples from patients diagnosed with metastatic prostate cancer (mean ± SEM = 21 ± 2.957 CTCs/mL, median = 21 CTCs/mL), demonstrating the potential clinical utility of this device. In the last two decades, circulating tumor cells (CTCs) have attracted a significant amount of attention for their potential use as a blood-based biomarker for a broad range of cancer-related clinical applications. CTCs are malignant cells that are shed from the primary and/or metastatic solid tumors and then infiltrate into the vascular and lymphatic systems; these cells play a fundamental role in the metastatic process of non-hematological cancers 1-3. Although the first report describing the existence of CTCs dates from 1869 4 , the heterogeneity and the

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Section: Resultsmentioning

confidence: 99%

Characterization of a novel automated microfiltration device for the efficient isolation and analysis of circulating tumor cells from clinical blood samples

León¹,

Soto-García²,

Araíz-Hernández³

et al. 2020

Sci Rep

Self Cite

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“…Up until now, most of the micro uidic chip-based CTCs platforms still rely on manual operations for cell screening and interpretation after immuno uorescent staining of target cells, which is time-consuming and causes inconsistent results. So far, only a limited number of automated CTCs microscope systems have been reported 21 . The automated CTCs scanning and identi cation system can complete the scanning of the target cells in a short time, which not only saves time and manpower, but also helps to establish standard processes.…”

Section: Discussionmentioning

confidence: 99%

A Novel Automatic Platform Based on Nanostructured Microfluidic Chip for Isolating and Identification of Circulating Tumor Cells

Jou

Chou

Chang

et al. 2021

Preprint

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Circulating tumor cell (CTC) is commonly used as biomarker for early detection, prognostication, decision making, and evaluation of the therapeutic efficacy in cancer treatment. Unfortunately, the poor reproducibility and limited sensitivity with the CTC detection have limited its potential impact on clinical management. A reliable automated CTC detection system is therefore needed. We have designed an automated microfluidic chip-based CTC detection system consisting of three main parts: V-BioChip, Cell RevealTM enrichment and staining system, and an automatic scanning and locating system. We hypothesized this novel system can reliably detect CTC from clinical specimens. SKOV3 ovarian cancer cell line was used first to test the reliability of our system. Ten healthy volunteers, 5 patients with benign ovarian tumors, and 8 patients with epithelial ovarian cancer (EOC) were recruited after obtaining written consents to validate the CTC capturing efficacy in the peripheral blood. The capture rates for spiking test in SKOV3 cells were as 48.3% and 89.6% using EpCAM antibody alone and a combined EpCAM antibody and N-cadherin antibody, respectively. The system was sensitive to detection of low cell count and showed a linear relationship with the cell counts in our test range. The sensitivity and specificity were 62.5% and 100% when CTC was used as a biomarker for EOC. Our results demonstrated that this automatic CTC platform has a high capture rate and is feasible for detection of CTCs in EOC.

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“…The AccuCyte-CyteFinder was able to identify equivalent or greater numbers of CTCs found by the CellSearch system [ 68 ]. Aguilar-Avelar et al (2019) described the design and construction of a fully automated high-throughput fluorescence microscope that enables the recognition, imaging, and classification of CTCs in a blood sample that were labeled by immunostaining [ 69 ]. The microscope hardware accurately discriminated CTCs among cells present in blood and the hardware efficiently captured light emitted from unstained cells while the fluorescence signals were used to automatically classify and precisely identify CTCs on a sample by thousands of background cells [ 69 ].…”

Section: Genetic and Chromosome Instabilitymentioning

confidence: 99%

Genomic Instability in Circulating Tumor Cells

Freitas

Gartner

Rangel‐Pozzo

et al. 2020

Cancers

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Circulating tumor cells (CTCs) can promote distant metastases and can be obtained through minimally invasive liquid biopsy for clinical assessment in cancer patients. Having both genomic heterogeneity and instability as common features, the genetic characterization of CTCs can serve as a powerful tool for a better understanding of the molecular changes occurring at tumor initiation and during tumor progression/metastasis. In this review, we will highlight recent advances in the detection and quantification of tumor cell heterogeneity and genomic instability in CTCs. We will focus on the contribution of chromosome instability studies to genetic heterogeneity in CTCs at the single-CTC level by discussing data from different cancer subtypes and their impact on diagnosis and precision medicine.

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High-Throughput Automated Microscopy of Circulating Tumor Cells

Cited by 16 publications

References 37 publications

Characterization of a novel automated microfiltration device for the efficient isolation and analysis of circulating tumor cells from clinical blood samples

Characterization of a novel automated microfiltration device for the efficient isolation and analysis of circulating tumor cells from clinical blood samples

A Novel Automatic Platform Based on Nanostructured Microfluidic Chip for Isolating and Identification of Circulating Tumor Cells

Genomic Instability in Circulating Tumor Cells

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