Identifying EGFR-Expressed Cells and Detecting EGFR Multi-Mutations at Single-Cell Level by Microfluidic Chip.

Article Source
Related Articles

Identifying EGFR-Expressed Cells and Detecting EGFR Multi-Mutations at Single-Cell Level by Microfluidic Chip.

Nanomicro Lett. 2018;10(1):16

Authors: Li R, Zhou M, Li J, Wang Z, Zhang W, Yue C, Ma Y, Peng H, Wei Z, Hu Z

Abstract
EGFR mutations companion diagnostics have been proved to be crucial for the efficacy of tyrosine kinase inhibitor targeted cancer therapies. To uncover multiple mutations occurred in minority of EGFR-mutated cells, which may be covered by the noises from majority of un-mutated cells, is currently becoming an urgent clinical requirement. Here we present the validation of a microfluidic-chip-based method for detecting EGFR multi-mutations at single-cell level. By trapping and immunofluorescently imaging single cells in specifically designed silicon microwells, the EGFR-expressed cells were easily identified. By in situ lysing single cells, the cell lysates of EGFR-expressed cells were retrieved without cross-contamination. Benefited from excluding the noise from cells without EGFR expression, the simple and cost-effective Sanger’s sequencing, but not the expensive deep sequencing of the whole cell population, was used to discover multi-mutations. We verified the new method with precisely discovering three most important EGFR drug-related mutations from a sample in which EGFR-mutated cells only account for a small percentage of whole cell population. The microfluidic chip is capable of discovering not only the existence of specific EGFR multi-mutations, but also other valuable single-cell-level information: on which specific cells the mutations occurred, or whether different mutations coexist on the same cells. This microfluidic chip constitutes a promising method to promote simple and cost-effective Sanger’s sequencing to be a routine test before performing targeted cancer therapy.

PMID: 30393665 [PubMed]

Leave a Reply

Your email address will not be published. Required fields are marked *

*


Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(http://api.facebook.com/restserver.php?method=links.getStats&format=json&urls=http%3A%2F%2Fwww.single-cell-analysis.com%2Fa-novel-dual-well-array-chip-for-efficiently-trapping-single-cell-in-large-isolated-micro-well-without-complicated-accessory-equipment%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(http://urls.api.twitter.com/1/urls/count.json?url=http%3A%2F%2Fwww.single-cell-analysis.com%2Fa-novel-dual-well-array-chip-for-efficiently-trapping-single-cell-in-large-isolated-micro-well-without-complicated-accessory-equipment%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(http://api.facebook.com/restserver.php?method=links.getStats&format=json&urls=http%3A%2F%2Fwww.single-cell-analysis.com%2Fspectrofluorimetric-quantification-of-antibiotic-drug-concentration-in-bacterial-cells-for-the-characterization-of-translocation-across-bacterial-membranes%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(http://urls.api.twitter.com/1/urls/count.json?url=http%3A%2F%2Fwww.single-cell-analysis.com%2Fspectrofluorimetric-quantification-of-antibiotic-drug-concentration-in-bacterial-cells-for-the-characterization-of-translocation-across-bacterial-membranes%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(http://api.facebook.com/restserver.php?method=links.getStats&format=json&urls=http%3A%2F%2Fwww.single-cell-analysis.com%2Fsingle-cell-analysis-of-kynurenine-and-system-l-amino-acid-transport-in-t-cells%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(http://urls.api.twitter.com/1/urls/count.json?url=http%3A%2F%2Fwww.single-cell-analysis.com%2Fsingle-cell-analysis-of-kynurenine-and-system-l-amino-acid-transport-in-t-cells%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(http://api.facebook.com/restserver.php?method=links.getStats&format=json&urls=http%3A%2F%2Fwww.single-cell-analysis.com%2Fa-microfluidic-device-for-massively-parallel-whole-lifespan-imaging-of-single-fission-yeast-cells%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(http://urls.api.twitter.com/1/urls/count.json?url=http%3A%2F%2Fwww.single-cell-analysis.com%2Fa-microfluidic-device-for-massively-parallel-whole-lifespan-imaging-of-single-fission-yeast-cells%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(http://api.facebook.com/restserver.php?method=links.getStats&format=json&urls=http%3A%2F%2Fwww.single-cell-analysis.com%2Fsingle-cell-mrna-seq-using-the-fluidigm-c1-system-and-integrated-fluidics-circuits%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(http://urls.api.twitter.com/1/urls/count.json?url=http%3A%2F%2Fwww.single-cell-analysis.com%2Fsingle-cell-mrna-seq-using-the-fluidigm-c1-system-and-integrated-fluidics-circuits%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(http://api.facebook.com/restserver.php?method=links.getStats&format=json&urls=http%3A%2F%2Fwww.single-cell-analysis.com%2Fdual-nozzle-microfluidic-droplet-generator%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 121

Warning: file_get_contents(): http:// wrapper is disabled in the server configuration by allow_url_fopen=0 in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179

Warning: file_get_contents(http://urls.api.twitter.com/1/urls/count.json?url=http%3A%2F%2Fwww.single-cell-analysis.com%2Fdual-nozzle-microfluidic-droplet-generator%2F): failed to open stream: no suitable wrapper could be found in /home/rnaseqbl/single-cell-analysis.com/wp-content/plugins/most-shared-posts/msp-fetch.php on line 179