Document Detail


Preparing sample chambers for single-molecule FRET.
MedLine Citation:
PMID:  23028078     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Single-molecule (sm) fluorescence detection is a powerful method for studying biological events without time and population averaging. Förster (fluorescence) resonance energy transfer (FRET) is a spectroscopic technique in which the efficiency of energy transfer from donor to acceptor molecules is used to determine distances between molecules in the 30-80 Å range. Structural changes in biological molecules or relative motion between two interacting molecules can be detected by a change in FRET. A variant of smFRET is based on total internal reflection (TIR) microscopy, which can be set up in two ways, either using an oil-immersion (objective-type) or a water-immersion (prism-type) lens. To study the conformational changes of individual molecules over extended time periods, molecules must be localized in space. This protocol describes the preparation of sample chambers with either bovine serum albumin (BSA)- or polyethylene glycol (PEG)-coated slides to which single molecules can be tethered for use in FRET studies.
Authors:
Chirlmin Joo; Taekjip Ha
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Publication Detail:
Type:  Journal Article     Date:  2012-10-01
Journal Detail:
Title:  Cold Spring Harbor protocols     Volume:  2012     ISSN:  1559-6095     ISO Abbreviation:  Cold Spring Harb Protoc     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-02     Completed Date:  2013-02-26     Revised Date:  2013-06-11    
Medline Journal Info:
Nlm Unique ID:  101524530     Medline TA:  Cold Spring Harb Protoc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1104-8     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
DNA / chemistry,  metabolism
Fluorescence Resonance Energy Transfer / methods*
Microscopy / methods*
Molecular Biology / methods
Nucleic Acid Conformation
Protein Conformation
Proteins / chemistry,  metabolism
RNA / chemistry,  metabolism
Specimen Handling / methods*
Grant Support
ID/Acronym/Agency:
R01 GM065367/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Proteins; 63231-63-0/RNA; 9007-49-2/DNA

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


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