Title: Global Analysis of Microscopic Fluorescence Lifetime Images Using Spectral Segmentation and a Digital Micromirror Spatial Illuminator
Authors: BEDNARKIEWICZ ArturWHELAN Maurice
Citation: JOURNAL OF BIOMEDICAL OPTICS vol. 13 no. 4 p. 041316-1 041316-13
Publisher: SPIE-INT SOCIETY OPTICAL ENGINEERING
Publication Year: 2008
JRC N°: JRC47645
ISSN: 1083-3668
URI: http://publications.jrc.ec.europa.eu/repository/handle/JRC47645
Type: Articles in Journals
Abstract: Fluorescence lifetime imaging FLIM is very demanding from a technical and computational perspective, and the output is usually a compromise between acquisition/processing time and data accuracy and precision. We present a new approach to acquisition, analysis, and reconstruction of microscopic FLIM images by employing a digital micromirror device DMD as a spatial illuminator. In the first step, the whole field fluorescence image is collected by a color charge-coupled device CCD camera. Further qualitative spectral analysis and sample segmentation are performed to spatially distinguish between spectrally different regions on the sample. Next, the fluorescence of the sample is excited segment by segment, and fluorescence lifetimes are acquired with a photon counting technique. FLIM image reconstruction is performed by either raster scanning the sample or by directly accessing specific regions of interest. The unique features of the DMD illuminator allow the rapid on-line measurement of global good initial parameters GIP, which are supplied to the first iteration of the fitting algorithm. As a consequence, a decrease of the computation time required to obtain a satisfactory quality-of-fit is achieved without compromising the accuracy and precision of the lifetime measurements
JRC Institute:Institute for Health and Consumer Protection

Files in This Item:
There are no files associated with this item.


Items in repository are protected by copyright, with all rights reserved, unless otherwise indicated.