Commercial high speed machines open new opportunities in high throughput flow cytometry (HTFC)

Ashcroft, R.G.; Lopez, Peter

doi:10.1016/s0022-1759(00)00219-2

Cited by 66 publications

(36 citation statements)

References 34 publications

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“…MACS is a selection technique that is capable of capturing a large number of target cells in parallel (4); however, the purity and recovery in MACS typically have large variances (5). In contrast, FACS relies upon serially screening each cell, yielding high performance in cell recovery and purity (6). However, because of the serial nature of its operation, FACS allows for a comparatively low throughput, typically in the range between 10 4 and 10 5 cells per second (7).…”

mentioning

confidence: 99%

Marker-specific sorting of rare cells using dielectrophoresis

Bessette²,

Qian³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

409

299

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Current techniques in high-speed cell sorting are limited by the inherent coupling among three competing parameters of performance: throughput, purity, and rare cell recovery. Microfluidics provides an alternate strategy to decouple these parameters through the use of arrayed devices that operate in parallel. To efficiently isolate rare cells from complex mixtures, an electrokinetic sorting methodology was developed that exploits dielectrophoresis (DEP) in microfluidic channels. In this approach, the dielectrophoretic amplitude response of rare target cells is modulated by labeling cells with particles that differ in polarization response. Cell mixtures were interrogated in the DEP-activated cell sorter in a continuous-flow manner, wherein the electric fields were engineered to achieve efficient separation between the dielectrophoretically labeled and unlabeled cells. To demonstrate the efficiency of marker-specific cell separation, DEP-activated cell sorting (DACS) was applied for affinity-based enrichment of rare bacteria expressing a specific surface marker from an excess of nontarget bacteria that do not express this marker. Rare target cells were enriched by >200-fold in a single round of sorting at a single-channel throughput of 10,000 cells per second. DACS offers the potential for automated, surface marker-specific cell sorting in a disposable format that is capable of simultaneously achieving high throughput, purity, and rare cell recovery.cell sorting ͉ microfluidics C ell sorters are capable of separating a heterogeneous suspension of particles into purified fractions and thus have become an indispensable tool in biology and medicine. Emerging applications of cell sorting technology span a broad spectrum of pharmaceutical and biomedical fields that range from cancer diagnostics to cell-based therapies (1-3). The most widely used methodologies for cell separation are magnetic-activated cell sorting (MACS) and fluorescence-activated cell sorting (FACS). MACS is a selection technique that is capable of capturing a large number of target cells in parallel (4); however, the purity and recovery in MACS typically have large variances (5). In contrast, FACS relies upon serially screening each cell, yielding high performance in cell recovery and purity (6). However, because of the serial nature of its operation, FACS allows for a comparatively low throughput, typically in the range between 10 4 and 10 5 cells per second (7). Regardless of the mechanism, the performance of cell separation is typically characterized by three metrics. ''Throughput'' gauges how many cell characterization and sorting operations can be executed per unit of time, ''purity'' is the fraction of the target cells in the collection vessel, and ''recovery'' is the fraction of the input target cells successfully sorted into the collection vessel. Demands placed on cell sorting technologies continue to increase, because cell sorting applications are expanding and biological questions are becoming more complex (7). For example, rare cell sort...

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mentioning

confidence: 99%

Marker-specific sorting of rare cells using dielectrophoresis

Bessette²,

Qian³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

409

299

View full text Add to dashboard Cite

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“…Αυ τό έχει ως αποτέλεσμα να αναγκάζει τα κύτταρα να δια τάσσονται το ένα κατόπιν του άλλου και τελικά να διέρ- χονται από το ρύγχος του υδροδυναμικού συστήματος με ταχύτητα 3.000 -10.000 κυττάρων/sec (Darzynkiewicz andCrissman 1990, Ashcroft andLopez 2000) (Εικόνα 1). Κα θώς τα κύτταρα διέρχονται από το ρύγχος της κυψέλης ροής, μια δέσμη φωτός LASER προσπίπτει κάθετα πάνω σε αυτά.…”

Section: περιγραφη της μεθοδουunclassified

Flow cytometry and its applications

Papadogiannakis

Kontos

Tamamidou

et al. 2017

J Hellenic Vet Med Soc

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Flow cytometry is an automated, multiparametric and quantitative method that counts and analyses the scattering and fluorescence signals produced as a single cell stream passes through a laser beam. This article describes the method along with fluorochromes used and cell labeling techniques. Additionally, details regarding the applications of flow cytometry are also given. Although flow cytometry is not widely used in veterinary science so far, its application in the near future will rise, since more necessary reagents for veterinary use will be in the market.

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“…After fluorescent labeling, cells can be identified by fluorescence signals one by one in a flow cytometer. [1][2][3][4][5][6][7] Similarly, cells labeled with magnetic particles can be sorted by a magnetic substance. 8 By labeling specific proteins or antibodies, cells of interest can be affixed on microscale devices while unwanted cells pass through.…”

Section: Introductionmentioning

confidence: 99%

A cell sorting and trapping microfluidic device with an interdigital channel

Qiao

et al. 2016

AIP Advances

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The growing interest in cell sorting and trapping is driving the demand for high performance technologies. Using labeling techniques or external forces, cells can be identified by a series of methods. However, all of these methods require complicated systems with expensive devices. Based on inherent differences in cellular morphology, cells can be sorted by specific structures in microfluidic devices. The weir filter is a basic and efficient cell sorting and trapping structure. However, in some existing weir devices, because of cell deformability and high flow velocity in gaps, trapped cells may become stuck or even pass through the gaps. Here, we designed and fabricated a microfluidic device with interdigital channels for cell sorting and trapping. The chip consisted of a sheet of silicone elastomer polydimethylsiloxane and a sheet of glass. A square-wave-like weir was designed in the middle of the channel, comprising the interdigital channels. The square-wave pattern extended the weir length by three times with the channel width remaining constant. Compared with a straight weir, this structure exhibited a notably higher trapping capacity. Interdigital channels provided more space to slow down the rate of the pressure decrease, which prevented the cells from becoming stuck in the gaps. Sorting a mixture K562 and blood cells to trap cells demonstrated the efficiency of the chip with the interdigital channel to sort and trap large and less deformable cells. With stable and efficient cell sorting and trapping abilities, the chip with an interdigital channel may be widely applied in scientific research fields.

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Commercial high speed machines open new opportunities in high throughput flow cytometry (HTFC)

Cited by 66 publications

References 34 publications

Marker-specific sorting of rare cells using dielectrophoresis

Marker-specific sorting of rare cells using dielectrophoresis

Flow cytometry and its applications

A cell sorting and trapping microfluidic device with an interdigital channel

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