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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...

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Detalles Bibliográficos
Autores principales: Purnapatra, Subhajit B., Bera, Sampa, Mondal, Partha Pratim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2012
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.
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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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