Method and device for isolating cells from heterogeneous solution using microfluidic trapping vortices

A microfluidic device and a technology for separating cells, which can be used in measuring devices, fluid controllers, and laboratory containers, and can solve problems such as useless capture of cells

Active Publication Date: 2013-08-21
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the creation of vortices for concentration or filtration assistance purposes is disclosed, these structures are not designed to selectively trap cells within

Method used

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  • Method and device for isolating cells from heterogeneous solution using microfluidic trapping vortices
  • Method and device for isolating cells from heterogeneous solution using microfluidic trapping vortices
  • Method and device for isolating cells from heterogeneous solution using microfluidic trapping vortices

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 - Enrichment of rare cancer cells from blood

[0060] image 3 The microfluidic device 10 was adapted to isolate and concentrate cancer cells (20 microns in diameter) from normal human blood cells (2 to 15 microns in diameter) to demonstrate size-based enrichment and concentration in a high-throughput manner. utility. For clinical diagnosis, the enrichment and concentration of cancer cells from blood is of particular importance, as circulating tumor cells (CTCs) can provide real-time information on patient pathology and monitoring of cancer treatment. Isolation of live CTCs from blood in a rapid, efficient and label-free manner remains a major technical challenge: CTCs are rare events with rates as low as one cell per billion blood cells. While current strategies focus on the investigation of CTCs for diagnosis, there is a significant need to collect larger sample volumes of live CTCs for research purposes. This requires higher throughput processing of la...

Embodiment 2

[0069] Example 2 - Cell labeling and solution replacement

[0070] The microfluidic device 10 is also used to efficiently label cells with specific molecular markers. In conventional centrifugation, a sample of cells is labeled with a specific marker through a series of labeling and washing steps. This involves incubating the cells with the labeling reagent in a centrifuge tube, concentrating the cells to a pellet in a benchtop centrifuge, removing the supernatant containing unbound labeling reagent by manual pipetting, and manually resuspending the cells in new medium. These manipulations were performed within the microfluidic device 10 by trapping the cells in a fluid vortex and then exposing the trapped coiled cells to a labeling reagent followed by a PBS wash. Labeled cells are then released into collection vials in small volumes by reducing the flow rate.

[0071] Figure 6A-6D respectively shows the interception ( Figure 6A ), first solution replacement ( Figure...

Embodiment 3

[0074] Example 3 - Sequential operation: enrichment of rare cells followed by labeling

[0075] use Figure 7 The microfluidic device 10 shown successfully performed the multiple sequential sample preparation steps enabled by the centrifuge (eg, entrapped fluorescent solution exchange, reaction, and wash). In this embodiment, the microfluidic device 10 includes three inlets 18', 18'', and 18'''. One inlet 18' is connected to a syringe pump 22 for delivering the cell sample. The second syringe pump 24 is used to deliver the fluorescent agent. The third syringe pump 26 is used to deliver washing solution (PBS). Size-based retention of cancer cells from blood, serial fluorescent labeling, and analysis of released cells were performed in Figure 8A-Figure 8D The order of interception, first solution replacement, reaction, and second solution replacement is shown. Cells were then washed Figure 9 , which shows fluorescent images of cell clusters that were sequentially tr...

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Abstract

A method of isolating cells includes providing a microfluidic device having at least one microfluidic channel coupled to an inlet and an outlet, the at least one microfluidic channel comprises at least one expansion region disposed along the length thereof. The at least one expansion region is an abrupt increase in a cross-sectional dimension of the at least one microfluidic channel configured to generate a vortex within the at least one expansion region in response to fluid flow. A solution containing a population of cells at least some of which have diameters = 10 [mu]m flows into the inlet. A portion of cells is trapped within vortex created within the at least one expansion region. The trapped cells may then released from the expansion region.

Description

[0001] related application [0002] This application claims priority to US Provisional Patent Application No. 61 / 382,840, filed September 14, 2010. Claim priority under 35 U.S.C. §119. The aforementioned patent applications are incorporated by reference as if fully set forth herein. technical field [0003] The field of the invention generally relates to microfluidic devices for separating and sorting cells or particles. More specifically, the field of the invention relates to microfluidic devices and methods that employ microfluidic entrapment vortices to separate cells or particles from heterogeneous solutions. Background technique [0004] Standard benchtop centrifuges are one of the most common instruments used in life science laboratories and are widely used for sample preparation in cell biology research and medical diagnostics. Typical sample preparation processes require multiple centrifugation steps for cell labeling and washing, which can be a time-consuming, la...

Claims

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Application Information

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IPC IPC(8): C12Q1/24G01N33/48G01N35/00
CPCB01L3/502746B01L3/502761B01L2200/0668B01L2300/0816B01L2300/0864B01L2300/0867B01L2300/087B01L2400/0487B01L2400/082B01L2400/086C12M47/04G01N1/40C12M23/16C12M29/00C12Q1/24C12Q1/68G01N1/4077G01N33/5091G01N33/582
Inventor 迪诺·迪卡洛许秀晶阿尔贝特·J·马赫
Owner RGT UNIV OF CALIFORNIA
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