Rare cell isolation and recovery on open-channel microfluidic chip

Masuda, Taisuke; Song, Woneui; Nakanishi, Hayao; Wu, Lei; Noor, Anas Mohd; Arai, Fumihito

doi:10.1371/journal.pone.0174937

Cited by 21 publications

(11 citation statements)

References 36 publications

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“…One of the unique advantages of this open hydrogel molded microfluidic device is the ability to selectively analyze cells that possess desirable features through a combination of imaging and selective deposition of unique reagents. For example, it may be possible to analyze only the drug-resistant cells from patient biopsy samples through time-lapse imaging, and then selectively analyze the small fraction of drug resistant cellular genomes . For proof of concept, we stained cells with a dye that fluoresces in the RFP channel and spiked them into the parental MDA-MB-231/GFP cell line at a 10:1 ratio.…”

Section: Resultsmentioning

confidence: 99%

Injection Molded Microfluidics for Establishing High-Density Single Cell Arrays in an Open Hydrogel Format

et al. 2020

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Here, we develop an injection molded microfluidic approach for single cell analysis by making use of (1) rapidly curing injectable hydrogels, (2) a high density microfluidic weir trap array, and (3) reversibly bonded PDMS lids that are strong enough to withstand the injection molding process, but which can be peeled off after the hydrogel sets. This approach allows for single cell patterns to be created with densities exceeding 40 cells per mm 2 , is amenable to high speed imaging, and creates microfluidic devices that enable efficient nutrient and gas exchange and the delivery of specific biological and chemical reagents to individual cells. We show that it is possible to organize up to 10 000 single cells in a few minutes on the device, and we developed an image analysis program to automatically analyze the single-cell capture efficiency. The results show single cell trapping rates were better than 80%. We also demonstrate that the genomic DNA of the single cells trapped in the hydrogel can be amplified via localized, multiple displacement amplification in a massively parallel format, which offers a promising strategy for analyzing single cell genomes. Finally, we show the ability to perform selective staining of individual cells with a commercial bioprinter, providing proof of concept of its ability to deliver tailored reagents to specific cells in an array for future downstream analysis. This injection molded microfluidic approach leverages the benefits of both closed and open microfluidics, allows multiday single cell cultures, direct access to the trapped cells for genotypic end point studies.

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Section: Resultsmentioning

confidence: 99%

Injection Molded Microfluidics for Establishing High-Density Single Cell Arrays in an Open Hydrogel Format

et al. 2020

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“…This separation method relies on cell size properties, as well as on a combination of size and deformability criteria. Several strategies were studied for CTC isolation: microstructure post filters [28][29][30][31][32], microporous membranes [33][34][35][36][37], or microfluidic constrictions [38][39][40][41][42][43].…”

Section: Microfiltration Separation Methodsmentioning

confidence: 99%

“…The first approach consists in integrating an array of micrometric-sized posts that act as filters to capture CTCs. To this end, several microstructure post shapes were imagined: from microellipse [30] to micropillars [31,32] and funnel constrictions [29]. Chen et al integrated an array of microellipse filters which consists of microfluidic slits in gradually narrowing series [30].…”

Section: Microfiltration Separation Methodsmentioning

confidence: 99%

Microfluidic-Based Technologies for CTC Isolation: A Review of 10 Years of Intense Efforts towards Liquid Biopsy

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Roy

Deman

2022

IJMS

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The selection of circulating tumor cells (CTCs) directly from blood as a real-time liquid biopsy has received increasing attention over the past ten years, and further analysis of these cells may greatly aid in both research and clinical applications. CTC analysis could advance understandings of metastatic cascade, tumor evolution, and patient heterogeneity, as well as drug resistance. Until now, the rarity and heterogeneity of CTCs have been technical challenges to their wider use in clinical studies, but microfluidic-based isolation technologies have emerged as promising tools to address these limitations. This review provides a detailed overview of latest and leading microfluidic devices implemented for CTC isolation. In particular, this study details must-have device performances and highlights the tradeoff between recovery and purity. Finally, the review gives a report of CTC potential clinical applications that can be conducted after CTC isolation. Widespread microfluidic devices, which aim to support liquid-biopsy-based applications, will represent a paradigm shift for cancer clinical care in the near future.

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“…We identified WBCs as CD45 and Hoechst + . We adopted this protocol to retrieve live cells without fixation and permeabilization, a process often used to label cytokeratin [26,21,22,32].…”

Section: Immunofluorescence Analysismentioning

confidence: 99%

High-purity isolation of rare single cells from blood using a tiered microchip system

et al. 2020

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We present a two-tiered microchip system to capture and retrieve rare cells from blood samples with high purity. The first module of the system is a high throughput microfluidic interface that is used to immunomagnetically isolate targeted rare cells from whole blood, and discard > 99.999% of the unwanted leukocytes. The second module is a microwell array that furthers the purification by magnetically guiding each cell into a separate well concurrently, and allows individual retrieval of each cell. We demonstrate the design of the system as well as its characterization by experiments using model cell lines that represent circulating fetal trophoblasts. Our results show that single cells can be retrieved with efficiencies and purities as high as 100% within 145 mins.

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Rare cell isolation and recovery on open-channel microfluidic chip

Cited by 21 publications

References 36 publications

Injection Molded Microfluidics for Establishing High-Density Single Cell Arrays in an Open Hydrogel Format

Injection Molded Microfluidics for Establishing High-Density Single Cell Arrays in an Open Hydrogel Format

Microfluidic-Based Technologies for CTC Isolation: A Review of 10 Years of Intense Efforts towards Liquid Biopsy

High-purity isolation of rare single cells from blood using a tiered microchip system

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