Improving fluorescence imaging of biological cells on biomedical polymers

Immunoﬂuorescence imaging on polymeric biomaterials is often inhibited by autoﬂuorescence and other optical phenomena. This often limits the analysis that can be performed on cells that are in contact with these materials. This study outlines a method that will quench these inhibitive optical phenom...

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Main Authors:	Jaafar, Israd Hakim, LeBlon, Courtney E., Wei, Ming-Tzo, Ou-Yang, Daniel, Coulter, John P., Jedlicka, Sabrina S.
Format:	Article
Language:	English
Published:	Elsevier 2011
Subjects:	TA164 Bioengineering
Online Access:	http://irep.iium.edu.my/153/ http://irep.iium.edu.my/153/ http://irep.iium.edu.my/153/ http://irep.iium.edu.my/153/1/Israd_Paper.pdf

id	iium-153
recordtype	eprints
spelling	iium-1532017-11-21T08:10:26Z http://irep.iium.edu.my/153/ Improving fluorescence imaging of biological cells on biomedical polymers Jaafar, Israd Hakim LeBlon, Courtney E. Wei, Ming-Tzo Ou-Yang, Daniel Coulter, John P. Jedlicka, Sabrina S. TA164 Bioengineering Immunoﬂuorescence imaging on polymeric biomaterials is often inhibited by autoﬂuorescence and other optical phenomena. This often limits the analysis that can be performed on cells that are in contact with these materials. This study outlines a method that will quench these inhibitive optical phenomena on a variety of polymeric materials, including poly(glycerol sebacate), poly(urethane), poly(L-lactide–co-ecaprolactone), and poly(lactic acid–co-glycolic acid). The method uses a simple material treatment method utilizing Sudan Black B (SB), which is commonly used as an autoﬂuorescence quenching molecule in tissue histology, but has not yet been used in biomaterials analysis. The quenching mechanism in the selected polymers is investigated using attenuated total reﬂectance Fourier transform infrared spectroscoy, ultraviolet–visible light absorbance and ﬂuorescence analysis, and scanning electron microscopyobservation of the material morphology prior to and after SB treatment. The results point to SB eliminating the inhibitive light phenomena of these materials by two methods: (i) chemical interaction between SB and the polymer molecules and (ii) physical interaction whereby SB forms a physical barrier that can absorb scattered light and quench autoﬂuorescence interference during ﬂuorescence microscopy. The studies show that the treatment of polymers with SB is robust across the polymers tested, in both porous and non-porous formats. The method does not interfere with immunoﬂuorescent imaging of ﬂuorescently labeled biological cells cultured on these polymers. This quick, simple, and affordable method enables a variety of analyses to be conducted that may otherwise have been impractical or impossible. Elsevier 2011 Article PeerReviewed application/pdf en http://irep.iium.edu.my/153/1/Israd_Paper.pdf Jaafar, Israd Hakim and LeBlon, Courtney E. and Wei, Ming-Tzo and Ou-Yang, Daniel and Coulter, John P. and Jedlicka, Sabrina S. (2011) Improving fluorescence imaging of biological cells on biomedical polymers. Acta Biomaterialia, 7 (4). pp. 1588-1598. ISSN 1742-7061 http://dx.doi.org/10.1016/j.actbio.2010.12.007 doi:10.1016/j.actbio.2010.12.007
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	TA164 Bioengineering
spellingShingle	TA164 Bioengineering Jaafar, Israd Hakim LeBlon, Courtney E. Wei, Ming-Tzo Ou-Yang, Daniel Coulter, John P. Jedlicka, Sabrina S. Improving fluorescence imaging of biological cells on biomedical polymers
description	Immunoﬂuorescence imaging on polymeric biomaterials is often inhibited by autoﬂuorescence and other optical phenomena. This often limits the analysis that can be performed on cells that are in contact with these materials. This study outlines a method that will quench these inhibitive optical phenomena on a variety of polymeric materials, including poly(glycerol sebacate), poly(urethane), poly(L-lactide–co-ecaprolactone), and poly(lactic acid–co-glycolic acid). The method uses a simple material treatment method utilizing Sudan Black B (SB), which is commonly used as an autoﬂuorescence quenching molecule in tissue histology, but has not yet been used in biomaterials analysis. The quenching mechanism in the selected polymers is investigated using attenuated total reﬂectance Fourier transform infrared spectroscoy, ultraviolet–visible light absorbance and ﬂuorescence analysis, and scanning electron microscopyobservation of the material morphology prior to and after SB treatment. The results point to SB eliminating the inhibitive light phenomena of these materials by two methods: (i) chemical interaction between SB and the polymer molecules and (ii) physical interaction whereby SB forms a physical barrier that can absorb scattered light and quench autoﬂuorescence interference during ﬂuorescence microscopy. The studies show that the treatment of polymers with SB is robust across the polymers tested, in both porous and non-porous formats. The method does not interfere with immunoﬂuorescent imaging of ﬂuorescently labeled biological cells cultured on these polymers. This quick, simple, and affordable method enables a variety of analyses to be conducted that may otherwise have been impractical or impossible.
format	Article
author	Jaafar, Israd Hakim LeBlon, Courtney E. Wei, Ming-Tzo Ou-Yang, Daniel Coulter, John P. Jedlicka, Sabrina S.
author_facet	Jaafar, Israd Hakim LeBlon, Courtney E. Wei, Ming-Tzo Ou-Yang, Daniel Coulter, John P. Jedlicka, Sabrina S.
author_sort	Jaafar, Israd Hakim
title	Improving fluorescence imaging of biological cells on biomedical polymers
title_short	Improving fluorescence imaging of biological cells on biomedical polymers
title_full	Improving fluorescence imaging of biological cells on biomedical polymers
title_fullStr	Improving fluorescence imaging of biological cells on biomedical polymers
title_full_unstemmed	Improving fluorescence imaging of biological cells on biomedical polymers
title_sort	improving fluorescence imaging of biological cells on biomedical polymers
publisher	Elsevier
publishDate	2011
url	http://irep.iium.edu.my/153/ http://irep.iium.edu.my/153/ http://irep.iium.edu.my/153/ http://irep.iium.edu.my/153/1/Israd_Paper.pdf
first_indexed	2023-09-18T20:07:06Z
last_indexed	2023-09-18T20:07:06Z
_version_	1777407231840485376

Improving fluorescence imaging of biological cells on biomedical polymers

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