Spectrally Resolved Fluorescence Lifetime Imaging Microscopy
Authors: Hanley, Quentin S.; Arndt-Jovin, Donna J.; Jovin, Thomas M.
Source: Applied Spectroscopy, Volume 56, Issue 2, Pages 50A-69A and 145-274 (February 2002) , pp. 155-166(12)
Publisher: Society for Applied Spectroscopy
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 ~50 (λ/Δλ) in the current arrangement and a wavelength range of ~430-750 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 ~1 ns in buffer to ~18 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.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
Document Type: Research article
DOI: http://dx.doi.org/10.1366/0003702021954610
Publication date: 2002-02-01
- The Society publishes the internationally recognized, peer reviewed journal, Applied Spectroscopy, which is available both in print and online. Subscriptions are included with membership or can be purchased by institutional or corporate organizations. Abstracts may be viewed free of charge. Previously published as Bulletin (Society for Applied Spectroscopy)
- Editorial Board
- Information for Authors
- Submit a Paper
- Subscribe to this Title
- Membership Information
- Request copyrighted SAS materials
- Spectroscopic Nomenclature
- Focus Compendium
- ingentaconnect is not responsible for the content or availability of external websites
- In this: publication
- By this: publisher
- In this Subject: Analytical Chemistry
- By this author: Hanley, Quentin S. ; Arndt-Jovin, Donna J. ; Jovin, Thomas M.

Shopping cart
Receive new issue alert
Get Permissions