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Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy
Monitoring and visualizing specimens at a large penetration depth is a challenge. At depths of hundreds of microns, several physical effects (such as, scattering, PSF distortion and noise) deteriorate the image quality and prohibit a detailed study of key biological phenomena. In this study, we use...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3457039/ https://www.ncbi.nlm.nih.gov/pubmed/23012646 http://dx.doi.org/10.1038/srep00692 |
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author | Purnapatra, Subhajit B. Bera, Sampa Mondal, Partha Pratim |
author_facet | Purnapatra, Subhajit B. Bera, Sampa Mondal, Partha Pratim |
author_sort | Purnapatra, Subhajit B. |
collection | PubMed |
description | Monitoring and visualizing specimens at a large penetration depth is a challenge. At depths of hundreds of microns, several physical effects (such as, scattering, PSF distortion and noise) deteriorate the image quality and prohibit a detailed study of key biological phenomena. In this study, we use a Bessel-like beam in-conjugation with an orthogonal detection system to achieve depth imaging. A Bessel-like penetrating diffractionless beam is generated by engineering the back-aperture of the excitation objective. The proposed excitation scheme allows continuous scanning by simply translating the detection PSF. This type of imaging system is beneficial for obtaining depth information from any desired specimen layer, including nano-particle tracking in thick tissue. As demonstrated by imaging the fluorescent polymer-tagged-CaCO(3) particles and yeast cells in a tissue-like gel-matrix, the system offers a penetration depth that extends up to 650 µm. This achievement will advance the field of fluorescence imaging and deep nano-particle tracking. |
format | Online Article Text |
id | pubmed-3457039 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-34570392012-09-25 Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy Purnapatra, Subhajit B. Bera, Sampa Mondal, Partha Pratim Sci Rep Article Monitoring and visualizing specimens at a large penetration depth is a challenge. At depths of hundreds of microns, several physical effects (such as, scattering, PSF distortion and noise) deteriorate the image quality and prohibit a detailed study of key biological phenomena. In this study, we use a Bessel-like beam in-conjugation with an orthogonal detection system to achieve depth imaging. A Bessel-like penetrating diffractionless beam is generated by engineering the back-aperture of the excitation objective. The proposed excitation scheme allows continuous scanning by simply translating the detection PSF. This type of imaging system is beneficial for obtaining depth information from any desired specimen layer, including nano-particle tracking in thick tissue. As demonstrated by imaging the fluorescent polymer-tagged-CaCO(3) particles and yeast cells in a tissue-like gel-matrix, the system offers a penetration depth that extends up to 650 µm. This achievement will advance the field of fluorescence imaging and deep nano-particle tracking. Nature Publishing Group 2012-09-25 /pmc/articles/PMC3457039/ /pubmed/23012646 http://dx.doi.org/10.1038/srep00692 Text en Copyright © 2012, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareALike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/ |
spellingShingle | Article Purnapatra, Subhajit B. Bera, Sampa Mondal, Partha Pratim Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy |
title | Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy |
title_full | Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy |
title_fullStr | Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy |
title_full_unstemmed | Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy |
title_short | Spatial Filter Based Bessel-Like Beam for Improved Penetration Depth Imaging in Fluorescence Microscopy |
title_sort | spatial filter based bessel-like beam for improved penetration depth imaging in fluorescence microscopy |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3457039/ https://www.ncbi.nlm.nih.gov/pubmed/23012646 http://dx.doi.org/10.1038/srep00692 |
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