Document Detail

Two-color two-photon fluorescence laser scanning microscopy.
MedLine Citation:
PMID:  19590940     Owner:  NLM     Status:  MEDLINE    
We present the first realization of a Two-Color Two-Photon Laser-Scanning Microscope (2c2pLSM) and UV fluorescence images of cells acquired with this technique. Fluorescence is induced by two-color two-photon absorption using the fundamental and the second harmonic of a Ti:Sa femtosecond laser. Simultaneous absorption of an 800 nm photon and a 400 nm photon energetically corresponds to one-photon absorption at 266 nm. This technique for Laser-Scanning Microscopy extends the excitation wavelength range of a Ti:Sa powered fluorescence microscope to the UV. In addition to the known advantages of multi-photon microscopy like intrinsic 3D resolution, reduced photo damage and high penetration depth 2c2pLSM offers the possibility of using standard high numeric aperture objectives for UV fluorescence imaging. The effective excitation wavelength of 266 nm corresponds especially well to the excitation spectrum of tryptophan. Hence, it is an ideal tool for label free fluorescence studies and imaging of intrinsic protein fluorescence which originates mainly from tryptophan. Thus a very sensitive natural lifetime probe can be used for monitoring protein reactions or changes in conformation. First measurements of living MIN-6 cells reveal differences between the UV fluorescence lifetimes of the nucleus and cytoplasm. The significance of this method was further demonstrated by monitoring the binding of biotin to avidin.
S Quentmeier; S Denicke; K-H Gericke
Publication Detail:
Type:  Journal Article     Date:  2009-07-10
Journal Detail:
Title:  Journal of fluorescence     Volume:  19     ISSN:  1573-4994     ISO Abbreviation:  J Fluoresc     Publication Date:  2009 Nov 
Date Detail:
Created Date:  2009-11-26     Completed Date:  2010-02-12     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9201341     Medline TA:  J Fluoresc     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1037-43     Citation Subset:  IM    
Institute for Physical und Theoretical Chemistry, University of Braunschweig, Hans-Sommer-Str. 10, 38106 Braunschweig, Germany.
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MeSH Terms
Avidin / chemistry
Biotin / chemistry
Cell Line, Tumor
Cell Nucleus / chemistry
Cytoplasm / chemistry
Microscopy, Confocal / instrumentation,  methods*
Microscopy, Fluorescence, Multiphoton / instrumentation,  methods*
Normal Distribution
Protein Binding
Ultraviolet Rays
Reg. No./Substance:
1405-69-2/Avidin; 58-85-5/Biotin

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

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