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Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy
The influence of chromatic aberration of an objective lens in two-photon fluorescence (TPF) endospectroscopy of scattering media has been systematically investigated through both experiments and numerical simulations. Experiments were carried out on a uniform 3D scattering gelatin phantom embedded w...
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Formato: | Texto |
Lenguaje: | English |
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Optical Society of America
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3018084/ https://www.ncbi.nlm.nih.gov/pubmed/21258545 http://dx.doi.org/10.1364/BOE.1.001234 |
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author | Wu, Yicong Li, Xingde |
author_facet | Wu, Yicong Li, Xingde |
author_sort | Wu, Yicong |
collection | PubMed |
description | The influence of chromatic aberration of an objective lens in two-photon fluorescence (TPF) endospectroscopy of scattering media has been systematically investigated through both experiments and numerical simulations. Experiments were carried out on a uniform 3D scattering gelatin phantom embedded with TiO(2) granules (to mimic tissue scattering) and fluorescein-tagged polystyrene beads. It was found that fluorescence spectral intensity and lineshape varied as a function of depth when measured with a gradient-index (GRIN) lens which has severe chromatic aberration. The spectral distortion caused by the chromatic aberration became diminishing as the imaging depth increased. Ray tracing analysis and Monte Carlo simulations were carried out to study the interplay of chromatic aberration and scattering in the depth-resolved TPF spectra. The simulation results suggest that the collected fluorescence signals from deeper layers included more out-of-focus photons that experienced a few or multiple scatterings, which diminish the influence of chromatic aberration on the measured TPF spectra. The simulated collection efficiencies of TPF at different wavelengths and depths can be used to properly recover the true depth-resolved TPF spectra of a relatively uniform scattering medium. |
format | Text |
id | pubmed-3018084 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Optical Society of America |
record_format | MEDLINE/PubMed |
spelling | pubmed-30180842011-01-21 Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy Wu, Yicong Li, Xingde Biomed Opt Express Microscopy The influence of chromatic aberration of an objective lens in two-photon fluorescence (TPF) endospectroscopy of scattering media has been systematically investigated through both experiments and numerical simulations. Experiments were carried out on a uniform 3D scattering gelatin phantom embedded with TiO(2) granules (to mimic tissue scattering) and fluorescein-tagged polystyrene beads. It was found that fluorescence spectral intensity and lineshape varied as a function of depth when measured with a gradient-index (GRIN) lens which has severe chromatic aberration. The spectral distortion caused by the chromatic aberration became diminishing as the imaging depth increased. Ray tracing analysis and Monte Carlo simulations were carried out to study the interplay of chromatic aberration and scattering in the depth-resolved TPF spectra. The simulation results suggest that the collected fluorescence signals from deeper layers included more out-of-focus photons that experienced a few or multiple scatterings, which diminish the influence of chromatic aberration on the measured TPF spectra. The simulated collection efficiencies of TPF at different wavelengths and depths can be used to properly recover the true depth-resolved TPF spectra of a relatively uniform scattering medium. Optical Society of America 2010-10-27 /pmc/articles/PMC3018084/ /pubmed/21258545 http://dx.doi.org/10.1364/BOE.1.001234 Text en ©2010 Optical Society of America http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 Unported License, which permits download and redistribution, provided that the original work is properly cited. This license restricts the article from being modified or used commercially. |
spellingShingle | Microscopy Wu, Yicong Li, Xingde Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy |
title | Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy |
title_full | Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy |
title_fullStr | Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy |
title_full_unstemmed | Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy |
title_short | Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy |
title_sort | combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy |
topic | Microscopy |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3018084/ https://www.ncbi.nlm.nih.gov/pubmed/21258545 http://dx.doi.org/10.1364/BOE.1.001234 |
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