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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my.iium.irep.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 REM 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 |
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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 |
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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 |
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Elsevier |
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2011 |
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http://irep.iium.edu.my/153/1/Israd_Paper.pdf http://irep.iium.edu.my/153/ http://dx.doi.org/10.1016/j.actbio.2010.12.007 |
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1643604544924418048 |
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13.19449 |