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In vivo quantification of fluorescent molecular markers in real‐time by ratio imaging for diagnostic screening and image‐guided surgery
Author(s) -
Bogaards A.,
Sterenborg H.J.C.M,
Trachtenberg J.,
Wilson B.C.,
Lilge L.
Publication year - 2007
Publication title -
lasers in surgery and medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.888
H-Index - 112
eISSN - 1096-9101
pISSN - 0196-8092
DOI - 10.1002/lsm.20525
Subject(s) - molecular imaging , autofluorescence , fluorescence , fluorescence lifetime imaging microscopy , preclinical imaging , optical imaging , biomedical engineering , in vivo , materials science , optics , medicine , physics , biology , microbiology and biotechnology
Future applications of “molecular diagnostic screening” and “molecular image‐guided surgery” will demand images of molecular markers with high resolution and high throughput (∼≥30 frames/second). MRI, SPECT, PET, optical fluorescence tomography, hyper‐spectral fluorescence imaging, and bioluminescence imaging do not offer such high frame rates. 2D optical fluorescence imaging can provide surface images with high resolution and high throughput. The ability to accurately quantify the fluorescence in vivo is critical to extract functional information of the disease state, however few methods are available. Here, a ratiometric 2D quantification method is introduced. Through mathematical modeling the performance was evaluated using optical properties that resembled biological tissues with the fluorescent marker Protoporhyrin IX. Experimentally the performance was evaluated in optical phantoms with different optical properties employing a novel prototype clinical imaging system. The clinical feasibility of real‐time, image‐guided surgery was demonstrated in patients undergoing prostatectomy. Discussed are the reasons why the introduced method leads to an increased quantification performance followed by modifications so it can be applied to novel fluorescent molecular markers as phthalocyanine 4 and dual‐fluorescent markers. These offer additional advantages as these can provide a linear response to marker concentration and further minimize the dependence on autofluorescence and optical properties, as demonstrated through modeling. Lesers Surg. Med. 39:605–613, 2007. © 2007 Wiley‐Liss, Inc.