Methods and devices for correlated, multi-parameter single cell measurements and recovery of remnant biological material

A single-cell and cell-based technology, applied in the direction of analytical materials, chemical instruments and methods, and laboratory containers, can solve problems such as unproduced methods and instruments

CN101842159AActive Publication Date: 2010-09-22CELULA CHINA MED TECH CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2010-09-22

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Abstract

Methods and apparatus are provided for analysis and correlation of phenotypic and genotypic information for a high throughput sample on a cell by cell basis. Cells are isolated and sequentially analyzed for phenotypic information and genotypic information which is then correlated. Methods for correlating the phenotype-genotype information of a sample population can be performed on a continuous flow sample within a microfluidic channel network or alternatively on a sample preloaded into a nano-well array chip. The methods for performing the phenotype-genotype analysis and correlation are scalable for samples numbering in the hundreds of cells to thousands of cells up to the tens and hundreds of thousand cells.
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Description

technical field

[0001] The present invention relates to a method and device for automatic and associated quantitative determination of protein and nucleic acid in a high-throughput sample obtained by sorting and collecting genomic materials cell by cell.

[0002] related application

[0003] This international application claims the benefit of Provisional Application No. 60 / 954,946, filed August 9, 2007, entitled "Associated Multiparametric Single-Cell Assays and Methods for Recovery of Residual Biological Material," which is incorporated by reference in its entirety into this article. Background technique

[0004] Fluorescence-activated cell sorting (FACS) is widely used in research and clinical applications. Such instruments have the ability to perform multiparameter analysis and sorting very quickly, but they typically require large sample volumes and trained operators for operation and maintenance, and are difficult to sterilize. FACS instruments can analyze as few a...

Examples

Embodiment Construction

[0042] The present invention provides methods suitable for cell-by-cell analysis of phenotypes (e.g., with fluorescently conjugated antibodies or components) for samples ranging in size from hundreds to thousands of cells to tens of thousands to hundreds of thousands of cells. Type-expressed fluorescent reporter molecules (such as green fluorescent protein) tagged cell surface proteins or intracellular proteins) information and genotype (such as for detection of single nucleotide polymorphisms (SNPs), DNA copy number or RNA gene expression Methods and devices for single cell, multiplex quantitative PCR or RT-PCR) information. Larger sample sizes facilitate the use of multidimensional scatterplots to reveal biological patterns and relationships between genes and proteins. Such as figure 1 As shown, the main purpose of the present invention is to obtain selected phenotypic characteristics and selected gene expression by interrogating individual cells on a cell-by-cell basis, a...