Improved Excitation Light Rejection Enhances Small-Animal Fluorescent Optical Imaging

Small-animal fluorescence-enhanced imaging involves the detection of weak fluorescent signals emanating from nanomolar to picomolar concentrations of exogenous or endogenously produced fluorophore concurrent with the rejection of an overwhelmingly large component of backscattered excitation light. T...

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Main Authors: Kildong Hwang, Jessica P. Houston, John C. Rasmussen, Amit Joshi, Shi Ke, Chun Li, Eva M. Sevick-Muraca
Format: Article
Language:English
Published: SAGE Publishing 2005-07-01
Series:Molecular Imaging
Online Access:https://doi.org/10.1162/15353500200505142
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author Kildong Hwang
Jessica P. Houston
John C. Rasmussen
Amit Joshi
Shi Ke
Chun Li
Eva M. Sevick-Muraca
author_facet Kildong Hwang
Jessica P. Houston
John C. Rasmussen
Amit Joshi
Shi Ke
Chun Li
Eva M. Sevick-Muraca
author_sort Kildong Hwang
collection DOAJ
description Small-animal fluorescence-enhanced imaging involves the detection of weak fluorescent signals emanating from nanomolar to picomolar concentrations of exogenous or endogenously produced fluorophore concurrent with the rejection of an overwhelmingly large component of backscattered excitation light. The elimination of the back-reflected excitation light of the collected signal remains a major and often unrecognized challenge for further reducing the noise floor and increasing sensitivity of small-animal fluorescence imaging. Herein, we show that the combination of three-cavity interference and holographic super notch filters with appropriate imaging lenses to collimate light improves rejection of excitation light, enabling more accurate imaging. To assess excitation leakage, the “out-of-band ( S (Λ x ))” to “in-band ( S (Λ m )– S (Λ x ))” signal ratio from phantom studies and the target-to-background ratio (TBR) from in vivo animal imaging was acquired with and without collimating optics. The addition of collimating optics resulted in a 51% to 75% reduction in the ratio of ( S (Λ x ))/( S (Λ m )– S (Λ x )) for the phantom studies and an improvement of TBR from 11% to 31% and of signal-to-noise ratio from 11% to 142% for an integrin-targeting conjugate in human glioma xenografts.
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institution Kabale University
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spelling doaj-art-b30446582c194cafb435c9c55668d7602025-02-03T10:07:39ZengSAGE PublishingMolecular Imaging1536-01212005-07-01410.1162/1535350020050514210.1162_15353500200505142Improved Excitation Light Rejection Enhances Small-Animal Fluorescent Optical ImagingKildong HwangJessica P. HoustonJohn C. RasmussenAmit JoshiShi KeChun LiEva M. Sevick-MuracaSmall-animal fluorescence-enhanced imaging involves the detection of weak fluorescent signals emanating from nanomolar to picomolar concentrations of exogenous or endogenously produced fluorophore concurrent with the rejection of an overwhelmingly large component of backscattered excitation light. The elimination of the back-reflected excitation light of the collected signal remains a major and often unrecognized challenge for further reducing the noise floor and increasing sensitivity of small-animal fluorescence imaging. Herein, we show that the combination of three-cavity interference and holographic super notch filters with appropriate imaging lenses to collimate light improves rejection of excitation light, enabling more accurate imaging. To assess excitation leakage, the “out-of-band ( S (Λ x ))” to “in-band ( S (Λ m )– S (Λ x ))” signal ratio from phantom studies and the target-to-background ratio (TBR) from in vivo animal imaging was acquired with and without collimating optics. The addition of collimating optics resulted in a 51% to 75% reduction in the ratio of ( S (Λ x ))/( S (Λ m )– S (Λ x )) for the phantom studies and an improvement of TBR from 11% to 31% and of signal-to-noise ratio from 11% to 142% for an integrin-targeting conjugate in human glioma xenografts.https://doi.org/10.1162/15353500200505142
spellingShingle Kildong Hwang
Jessica P. Houston
John C. Rasmussen
Amit Joshi
Shi Ke
Chun Li
Eva M. Sevick-Muraca
Improved Excitation Light Rejection Enhances Small-Animal Fluorescent Optical Imaging
Molecular Imaging
title Improved Excitation Light Rejection Enhances Small-Animal Fluorescent Optical Imaging
title_full Improved Excitation Light Rejection Enhances Small-Animal Fluorescent Optical Imaging
title_fullStr Improved Excitation Light Rejection Enhances Small-Animal Fluorescent Optical Imaging
title_full_unstemmed Improved Excitation Light Rejection Enhances Small-Animal Fluorescent Optical Imaging
title_short Improved Excitation Light Rejection Enhances Small-Animal Fluorescent Optical Imaging
title_sort improved excitation light rejection enhances small animal fluorescent optical imaging
url https://doi.org/10.1162/15353500200505142
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AT amitjoshi improvedexcitationlightrejectionenhancessmallanimalfluorescentopticalimaging
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