Document Detail


Selective encapsulation of single cells and subcellular organelles into picoliter- and femtoliter-volume droplets.
MedLine Citation:
PMID:  15762555     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This paper describes a method, which combines optical trapping and microfluidic-based droplet generation, for selectively and controllably encapsulating a single target cell or subcellular structure, such as a mitochondrion, into a picoliter- or femtoliter-volume aqueous droplet that is surrounded by an immiscible phase. Once the selected cell or organelle is encased within the droplet, it is stably confined in the droplet and cannot be removed. We demonstrate in droplet the rapid laser photolysis of the single cell, which essentially "freezes" the state that the cell was in at the moment of photolysis and confines the lysate within the small volume of the droplet. Using fluorescein di-beta-d-galactopyranoside, which is a fluorogenic substrate for the intracellular enzyme beta-galactosidase, we also assayed the activity of this enzyme from a single cell following the laser-induced lysis of the cell. This ability to entrap individual selected cells or subcellular organelles should open new possibilities for carrying out single-cell studies and single-organelle measurements.
Authors:
Mingyan He; J Scott Edgar; Gavin D M Jeffries; Robert M Lorenz; J Patrick Shelby; Daniel T Chiu
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Analytical chemistry     Volume:  77     ISSN:  0003-2700     ISO Abbreviation:  Anal. Chem.     Publication Date:  2005 Mar 
Date Detail:
Created Date:  2005-03-14     Completed Date:  2007-01-24     Revised Date:  2007-12-03    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1539-44     Citation Subset:  IM    
Affiliation:
Department of Chemistry, University of Washington, Seattle, Washington 98195-1700, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Mast Cells* / enzymology
Mice
Microfluidics / methods*
Mitochondria* / enzymology
Photolysis
beta-Galactosidase / metabolism
Grant Support
ID/Acronym/Agency:
GM65293-01/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
EC 3.2.1.23/beta-Galactosidase

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


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