Document Detail


A novel homogeneous bioluminescence resonance energy transfer element for biomolecular detection with CCD camera or CMOS device.
MedLine Citation:
PMID:  18167056     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A novel optical signal element based on homogeneous bioluminescence resonance energy transfer (BRET) was developed for biomolecular detection. A fluorescent dye and alkaline phosphatase (AP) conjugate was used as a reporter and light-generation element for imaging detection platforms that use a CCD camera or CMOS chip-based devices. In the presence of a luminescence substrate, the energy from the first light emission of a bioluminescence enzymatic reaction was transferred to fluorescent dyes which were conjugated to an enzyme. This resulted in a second light emission with a shorter wavelength. The second light was localized at the position of target molecules without the diffusion problems present in current technology. To optimize energy transfer efficiency, the ratio of enzyme to fluorophore in the conjugates, the fluorescent dyes used in the conjugates and the luminescence substrates used for BRET were investigated. BRET was demonstrated by using both a CCD camera and a CMOS imaging device. Image spatial resolution was greatly improved compared with conventional chemiluminescence detection. This new signal element opens a door for the direct measurement of fluorescent signals on an imaging chip without an external light source and portable instrumentation normally required for the fluorescent detection of biomolecules.
Authors:
Brian Filanoski; Shiva K Rastogi; Eric Cameron; Nirankar N Mishra; Wusi Maki; Gary Maki
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Luminescence : the journal of biological and chemical luminescence     Volume:  23     ISSN:  1522-7243     ISO Abbreviation:  Luminescence     Publication Date:    2008 Jan-Feb
Date Detail:
Created Date:  2008-01-23     Completed Date:  2008-04-18     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100889025     Medline TA:  Luminescence     Country:  England    
Other Details:
Languages:  eng     Pagination:  22-7     Citation Subset:  IM    
Copyright Information:
(c) 2007 John Wiley & Sons, Ltd.
Affiliation:
Center for Advanced Microelectronics and Biomolecular Research, University of Idaho, Post Falls, ID 83854, USA.
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MeSH Terms
Descriptor/Qualifier:
Alkaline Phosphatase / chemistry*
Energy Transfer
Fluorescent Dyes / chemistry*
Luminescence
Luminescent Measurements / methods*
Sensitivity and Specificity
Signal Processing, Computer-Assisted / instrumentation*
Time Factors
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; EC 3.1.3.1/Alkaline Phosphatase

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


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