English
 
User Manual Privacy Policy Disclaimer Contact us
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT
  Spectrally resolved fluorescence lifetime imaging microscopy

Hanley, Q. S., Arndt-Jovin, D. J., & Jovin, T. M. (2002). Spectrally resolved fluorescence lifetime imaging microscopy. Applied Spectroscopy, 56(2), 155-166. Retrieved from http://www.opticsinfobase.org/viewmedia.cfm?uri=as-56-2-155&seq=0.

Item is

Basic

show hide
Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0012-F45D-7 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-E5B5-8
Genre: Journal Article

Files

show Files

Locators

show

Creators

show
hide
 Creators:
Hanley, Q. S.1, Author              
Arndt-Jovin, D. J.1, Author              
Jovin, T. M.1, Author              
Affiliations:
1Department of Molecular Biology, MPI for biophysical chemistry, Max Planck Society, ou_578628              

Content

show
hide
Free keywords: S matrixes; LCD; spatial encoding; imaging spectroscopy; spectral imaging; microscopy; Hadamard transforms; fluorescence lifetimes; EGFP; propidium iodide; fluorescent proteins; frequency domain; global analysis; fluorescence lifetime imaging microscopy; FLIM; fluorescence resonance energy transfer; FRET
 Abstract: We report a system for collecting spectrally resolved fluorescent lifetime images. Frequency domain fluorescence lifetime detection was combined with two-dimensional spectral imaging in a programmable array microscope. The spectroscopic fluorescence lifetime imaging microscopy (sFLIM) system has a resolution of similar to50 (lambda/Deltalambda) in the current arrangement and a wavelength range of similar to430-750 nm. With the sFLIM system, we recorded the lifetime spectra of rhodamine 6G, rhodamine B, and the DNA intercalation dye propidium iodide (PI) in cuvettes and an EGFP-fusion of the histone 2A variant D protein in Drosophila salivary gland explants in the presence and absence of PI. In the absence of PI, the EGFP-fusion exhibited a lifetime of 2.7 ns with little variation in wavelength. The lifetime of PI alone ranged from similar to1 ns in buffer to similar to18 ns when intercalated in the nuclei of intact cells. The combination of EGFP and PI in the Drosophila salivary gland explants exhibited strong fluorescence resonance energy transfer (FRET), a result consistent with the known nucleosomal structure of eukaryotic chromatin.

Details

show
hide
Language(s): eng - English
 Dates: 2002-02
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Degree: -

Event

show

Legal Case

show

Project information

show

Source 1

show
hide
Title: Applied Spectroscopy
Source Genre: Journal
 Creator(s):
Affiliations:
Publ. Info: -
Pages: - Volume / Issue: 56 (2) Sequence Number: - Start / End Page: 155 - 166 Identifier: -