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A Simple low-cost device enables four epi-illumination techniques on standard light microscopes

Back-scattering darkfield (BSDF), epi-fluorescence (EF), interference reflection contrast (IRC), and darkfield surface reflection (DFSR) are advanced but expensive light microscopy techniques with limited availability. Here we show a simple optical design that combines these four techniques in a sim...

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Autores principales: Ishmukhametov, Robert R., Russell, Aidan N., Wheeler, Richard J., Nord, Ashley L., Berry, Richard M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4745110/
https://www.ncbi.nlm.nih.gov/pubmed/26853732
http://dx.doi.org/10.1038/srep20729
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author Ishmukhametov, Robert R.
Russell, Aidan N.
Wheeler, Richard J.
Nord, Ashley L.
Berry, Richard M.
author_facet Ishmukhametov, Robert R.
Russell, Aidan N.
Wheeler, Richard J.
Nord, Ashley L.
Berry, Richard M.
author_sort Ishmukhametov, Robert R.
collection PubMed
description Back-scattering darkfield (BSDF), epi-fluorescence (EF), interference reflection contrast (IRC), and darkfield surface reflection (DFSR) are advanced but expensive light microscopy techniques with limited availability. Here we show a simple optical design that combines these four techniques in a simple low-cost miniature epi-illuminator, which inserts into the differential interference-contrast (DIC) slider bay of a commercial microscope, without further additions required. We demonstrate with this device: 1) BSDF-based detection of Malarial parasites inside unstained human erythrocytes; 2) EF imaging with and without dichroic components, including detection of DAPI-stained Leishmania parasite without using excitation or emission filters; 3) RIC of black lipid membranes and other thin films, and 4) DFSR of patterned opaque and transparent surfaces. We believe that our design can expand the functionality of commercial bright field microscopes, provide easy field detection of parasites and be of interest to many users of light microscopy.
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spelling pubmed-47451102016-02-16 A Simple low-cost device enables four epi-illumination techniques on standard light microscopes Ishmukhametov, Robert R. Russell, Aidan N. Wheeler, Richard J. Nord, Ashley L. Berry, Richard M. Sci Rep Article Back-scattering darkfield (BSDF), epi-fluorescence (EF), interference reflection contrast (IRC), and darkfield surface reflection (DFSR) are advanced but expensive light microscopy techniques with limited availability. Here we show a simple optical design that combines these four techniques in a simple low-cost miniature epi-illuminator, which inserts into the differential interference-contrast (DIC) slider bay of a commercial microscope, without further additions required. We demonstrate with this device: 1) BSDF-based detection of Malarial parasites inside unstained human erythrocytes; 2) EF imaging with and without dichroic components, including detection of DAPI-stained Leishmania parasite without using excitation or emission filters; 3) RIC of black lipid membranes and other thin films, and 4) DFSR of patterned opaque and transparent surfaces. We believe that our design can expand the functionality of commercial bright field microscopes, provide easy field detection of parasites and be of interest to many users of light microscopy. Nature Publishing Group 2016-02-08 /pmc/articles/PMC4745110/ /pubmed/26853732 http://dx.doi.org/10.1038/srep20729 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Ishmukhametov, Robert R.
Russell, Aidan N.
Wheeler, Richard J.
Nord, Ashley L.
Berry, Richard M.
A Simple low-cost device enables four epi-illumination techniques on standard light microscopes
title A Simple low-cost device enables four epi-illumination techniques on standard light microscopes
title_full A Simple low-cost device enables four epi-illumination techniques on standard light microscopes
title_fullStr A Simple low-cost device enables four epi-illumination techniques on standard light microscopes
title_full_unstemmed A Simple low-cost device enables four epi-illumination techniques on standard light microscopes
title_short A Simple low-cost device enables four epi-illumination techniques on standard light microscopes
title_sort simple low-cost device enables four epi-illumination techniques on standard light microscopes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4745110/
https://www.ncbi.nlm.nih.gov/pubmed/26853732
http://dx.doi.org/10.1038/srep20729
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