Document Detail


Intracellular Tracking of Single Native Molecules with Electroporation-delivered Quantum Dots.
MedLine Citation:
PMID:  25341054     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Quantum dots (QDs) have found a wide range of biological applications as fluorophores due to their extraordinary brightness and high photostability that are far superior to conventional organic dyes. These traits are particularly appealing for studying cell biology under cellular autofluorescence background and with long observation period. However, it remains the most important open challenge to target QDs at native intracellular molecules and organelles in live cells. Endocytosis-based delivery methods lead to QDs encapsulated in vesicles that have their surface biorecognition element hidden from the intracellular environment. The probing of native molecules using QDs has been seriously hindered by the lack of consistent approaches for delivery of QDs with exposed surface groups. In this report, we demonstrate that electroporation (i.e. the application of short electric pulses for cell permeabilization) generates reproducible results for delivering QDs into cells. We show evidence that electroporation-based delivery does not involve endocytosis or vesicle encapsulation of QDs. The amount of QD loading and the resulted cell viability can be adjusted by varying the parameters associated with the electroporation operation. To demonstrate the application of our approach for intracellular targeting, we study single molecule motility of kinesin in live cells by labeling native kinesins using electroporation-delivered QDs. We envision that electroporation may serve as a simple and universal tool for delivering QDs into cells to label and probe native molecules and organelles.
Authors:
Chen Sun; Zhenning Cao; Min Wu; Chang Lu
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2014-10-23
Journal Detail:
Title:  Analytical chemistry     Volume:  -     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2014 Oct 
Date Detail:
Created Date:  2014-10-23     Completed Date:  -     Revised Date:  2014-10-24    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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