Document Detail


Microfluidic extraction, stretching and analysis of human chromosomal DNA from single cells.
MedLine Citation:
PMID:  23018789     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We describe a microfluidic device for the extraction, purification and stretching of human chromosomal DNA from single cells. A two-dimensional array of micropillars in a microfluidic polydimethylsiloxane channel was designed to capture a single human cell. Megabase-long DNA strands released from the cell upon lysis are trapped in the micropillar array and stretched under optimal hydrodynamic flow conditions. Intact chromosomal DNA is entangled in the array, while other cellular components are washed from the channel. To demonstrate the entrapment principle, a single chromosome was hybridized to whole chromosome paints, and imaged by fluorescence microscopy. DNA extracted from a single cell and small cell populations (less than 100) was released from the device by restriction endonuclease digestion under continuous flow and collected for off-chip analysis. Quantification of the extracted material reveals that the microdevice efficiently extracts essentially all chromosomal DNA. The device described represents a novel platform to perform a variety of analyses on chromosomal DNA at the single cell level.
Authors:
Jaime J Benítez; Juraj Topolancik; Harvey C Tian; Christopher B Wallin; David R Latulippe; Kylan Szeto; Patrick J Murphy; Benjamin R Cipriany; Stephen L Levy; Paul D Soloway; Harold G Craighead
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Lab on a chip     Volume:  12     ISSN:  1473-0189     ISO Abbreviation:  Lab Chip     Publication Date:  2012 Nov 
Date Detail:
Created Date:  2012-10-17     Completed Date:  2013-03-14     Revised Date:  2014-03-27    
Medline Journal Info:
Nlm Unique ID:  101128948     Medline TA:  Lab Chip     Country:  England    
Other Details:
Languages:  eng     Pagination:  4848-54     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Cell Line, Tumor
Chemical Fractionation / instrumentation*
Chromosomes, Human / genetics*
DNA / analysis*,  chemistry,  isolation & purification*
Humans
Microfluidic Analytical Techniques / instrumentation*
Nucleic Acid Conformation
Nucleic Acid Hybridization
Single-Cell Analysis / instrumentation*
Grant Support
ID/Acronym/Agency:
R01 DA025722/DA/NIDA NIH HHS; U54CA143876/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
9007-49-2/DNA
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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