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Microfluidic imaging cytometry

A technology of microfluidic system and microfluidic chip, which is applied in the field of microfluidic system and can solve problems such as no exploration platform

Inactive Publication Date: 2011-04-13
RGT UNIV OF CALIFORNIA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, no such exploration platform is currently available

Method used

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  • Microfluidic imaging cytometry
  • Microfluidic imaging cytometry
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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0098] 1. Cell culture sample preparation, including cell loading, culturing cells in an incubator, and media exchange for maintaining cells.

[0099] 2. Immunocytochemistry: including cell fixation, permeabilization and immunostaining.

[0100] According to some embodiments of the present invention, microfluidic system 100 may provide a system for (i) large-scale cell culture for high-throughput screening, (ii) microfluidic cell analysis for precise quantification of single-cell biomolecules , (iii) signaling pathway network analysis associated with cancer diagnosis and therapeutic classification, (iv) dynamic protein quantification as an alternative to Western blotting, and other broader applications for quantitative analysis of proteomics in cells.

[0101] Potential applications for the microfluidic system 100 may include, but are not limited to:

[0102] 1. Alternative techniques to flow cytometry, which may include the following advantages: (i) low cost, (ii) less patie...

Embodiment A

[0119] Example A: Determination Example

[0120] 1) Sample preparation, including cell loading, culturing cells in an incubator, and media exchange for maintaining cells.

[0121] 2) Immunocytochemistry: including cell fixation, permeabilization and immunostaining.

[0122] Can measure various signaling nodes (including EGFR, EGFvIII, PTEN, pAkt, pmTOR, pS6) involved in the PI3K-Akt-mTOR signaling network in the glioblastoma system and the proliferation marker Ki67.

[0123] Can measure various signaling nodes (including EGFR, ErbB2, PTEN, pAkt, pmTOR, pS6) and proliferation marker Ki67 involved in the PI3K-Akt-mTOR signaling network in the breast cancer system.

[0124] The growth curve of live cells can be monitored as follows:

[0125] The cell culture / assay chip shown in Figure 8 consists of two types of microchannels responsible for (i) parallel manipulation of 72 cell cultures and assays and (ii) wetting of adjacent cell culture chambers (to prevent media evaporation)....

Embodiment B

[0127] Example B: Data Collection

[0128] The data obtained by the methods of some embodiments of the invention may take the form of, for example, two-dimensional or three-dimensional dot plots (X-axis - strength of one signaling node (EGFRvIII in this case), Y-axis - another signaling Intensity of the node (DAPI in this case), for a 3D dot plot, Z-axis - intensity of the third signaling node))( Figure 9 ). In another embodiment ( Figure 10 ), the data can be in the form of a histogram (X-axis - strength of one signaling node (EGFRvIII in this case) and Y-axis - cell number).

[0129] Procedures for culturing glioblastoma cells in a chip

[0130] Material:

[0131] 24-channel poly-L-lysine coated cell culture chip

[0132] Cell culture medium: 500mL Dulbecco's Modified Eagle medium

[0133] 50mL fetal bovine serum

[0134] 5mL penicillin-streptomycin / L-glutamine

[0135] Cell lines and isogenetic cells:

[0136] U87

[0137] U87-PTEN

[0138] U87-EGFR

[0139] U...

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Abstract

A microfluidic system has a pipette system comprising a plurality of pipettes, a microfluidic chip arranged proximate the pipette system, an imaging optical detection system arranged proximate the microfluidic chip, and an image processing system in communication with the imaging optical detection system. The microfluidic chip has a plurality of cell culture chambers defined by a body of the microfluidic chip, each cell culture chamber being in fluid connection with an input channel and an output channel defined by the microfluidic chip. The pipette system is constructed and arranged to at least one of inject fluid through the plurality of pipettes into the plurality of input channels or extract fluid through the plurality of pipettes from the plurality of output channels while the microfluidic system is in operation.

Description

[0001] This application claims priority to US Provisional Patent Application no. 61 / 006842, filed February 1, 2008, the entire contents of which are incorporated herein by reference. technical field [0002] Embodiments of the present invention relate to microfluidic systems, and more particularly to microfluidic systems and methods for large scale cell culture and assays. Background technique [0003] All references cited in this specification are hereby incorporated by reference. [0004] Brain Tumor Classification Problems [0005] Astrocytic brain tumors comprise a diverse group of neoplasms with unique clinical, histopathological, and genetic features. Since the previous WHO classification in 1993, molecular genetic data collected have shown that the histologically defined types of astrocytomas are more distinct at the biological level 1 . For example, most glioblastomas develop without clinical or histological signs of less malignant premalignant lesions, which have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00G01N1/30
CPCG01N2035/00158G01N15/1484B01L2300/0816G01N2035/1039B01L2200/027C12M23/16G01N2015/1477B01L3/502715G01N2015/1006G01N35/1065B01L3/021
Inventor 曾宪荣龟井谦一郎孙晶保罗·S·米舍尔迈克尔·D·马斯特曼-史密斯大卫·A·内桑森黄亭漪迈克尔·万达姆克里斯蒂安·贝伦布鲁赫肖恩·M·萨尔卡里亚
Owner RGT UNIV OF CALIFORNIA
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