Cargando…
Combined Optical Coherence and Fluorescence Microscopy to assess dynamics and specificity of pancreatic beta-cell tracers
The identification of a beta-cell tracer is a major quest in diabetes research. However, since MRI, PET and SPECT cannot resolve individual islets, optical techniques are required to assess the specificity of these tracers. We propose to combine Optical Coherence Microscopy (OCM) with fluorescence d...
Autores principales: | , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4437378/ https://www.ncbi.nlm.nih.gov/pubmed/25988507 http://dx.doi.org/10.1038/srep10385 |
_version_ | 1782372202518151168 |
---|---|
author | Berclaz, Corinne Pache, Christophe Bouwens, Arno Szlag, Daniel Lopez, Antonio Joosten, Lieke Ekim, Selen Brom, Maarten Gotthardt, Martin Grapin-Botton, Anne Lasser, Theo |
author_facet | Berclaz, Corinne Pache, Christophe Bouwens, Arno Szlag, Daniel Lopez, Antonio Joosten, Lieke Ekim, Selen Brom, Maarten Gotthardt, Martin Grapin-Botton, Anne Lasser, Theo |
author_sort | Berclaz, Corinne |
collection | PubMed |
description | The identification of a beta-cell tracer is a major quest in diabetes research. However, since MRI, PET and SPECT cannot resolve individual islets, optical techniques are required to assess the specificity of these tracers. We propose to combine Optical Coherence Microscopy (OCM) with fluorescence detection in a single optical platform to facilitate these initial screening steps from cell culture up to living rodents. OCM can image islets and vascularization without any labeling. Thereby, it alleviates the need of both genetically modified mice to detect islets and injection of external dye to reveal vascularization. We characterized Cy5.5-exendin-3, an agonist of glucagon-like peptide 1 receptor (GLP1R), for which other imaging modalities have been used and can serve as a reference. Cultured cells transfected with GLP1R and incubated with Cy5.5-exendin-3 show full tracer internalization. We determined that a dose of 1 μg of Cy5.5-exendin-3 is sufficient to optically detect in vivo the tracer in islets with a high specificity. In a next step, time-lapse OCM imaging was used to monitor the rapid and specific tracer accumulation in murine islets and its persistence over hours. This optical platform represents a versatile toolbox for selecting beta-cell specific markers for diabetes research and future clinical diagnosis. |
format | Online Article Text |
id | pubmed-4437378 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-44373782015-06-01 Combined Optical Coherence and Fluorescence Microscopy to assess dynamics and specificity of pancreatic beta-cell tracers Berclaz, Corinne Pache, Christophe Bouwens, Arno Szlag, Daniel Lopez, Antonio Joosten, Lieke Ekim, Selen Brom, Maarten Gotthardt, Martin Grapin-Botton, Anne Lasser, Theo Sci Rep Article The identification of a beta-cell tracer is a major quest in diabetes research. However, since MRI, PET and SPECT cannot resolve individual islets, optical techniques are required to assess the specificity of these tracers. We propose to combine Optical Coherence Microscopy (OCM) with fluorescence detection in a single optical platform to facilitate these initial screening steps from cell culture up to living rodents. OCM can image islets and vascularization without any labeling. Thereby, it alleviates the need of both genetically modified mice to detect islets and injection of external dye to reveal vascularization. We characterized Cy5.5-exendin-3, an agonist of glucagon-like peptide 1 receptor (GLP1R), for which other imaging modalities have been used and can serve as a reference. Cultured cells transfected with GLP1R and incubated with Cy5.5-exendin-3 show full tracer internalization. We determined that a dose of 1 μg of Cy5.5-exendin-3 is sufficient to optically detect in vivo the tracer in islets with a high specificity. In a next step, time-lapse OCM imaging was used to monitor the rapid and specific tracer accumulation in murine islets and its persistence over hours. This optical platform represents a versatile toolbox for selecting beta-cell specific markers for diabetes research and future clinical diagnosis. Nature Publishing Group 2015-05-19 /pmc/articles/PMC4437378/ /pubmed/25988507 http://dx.doi.org/10.1038/srep10385 Text en Copyright © 2015, 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 Berclaz, Corinne Pache, Christophe Bouwens, Arno Szlag, Daniel Lopez, Antonio Joosten, Lieke Ekim, Selen Brom, Maarten Gotthardt, Martin Grapin-Botton, Anne Lasser, Theo Combined Optical Coherence and Fluorescence Microscopy to assess dynamics and specificity of pancreatic beta-cell tracers |
title | Combined Optical Coherence and Fluorescence Microscopy to assess dynamics and specificity of pancreatic beta-cell tracers |
title_full | Combined Optical Coherence and Fluorescence Microscopy to assess dynamics and specificity of pancreatic beta-cell tracers |
title_fullStr | Combined Optical Coherence and Fluorescence Microscopy to assess dynamics and specificity of pancreatic beta-cell tracers |
title_full_unstemmed | Combined Optical Coherence and Fluorescence Microscopy to assess dynamics and specificity of pancreatic beta-cell tracers |
title_short | Combined Optical Coherence and Fluorescence Microscopy to assess dynamics and specificity of pancreatic beta-cell tracers |
title_sort | combined optical coherence and fluorescence microscopy to assess dynamics and specificity of pancreatic beta-cell tracers |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4437378/ https://www.ncbi.nlm.nih.gov/pubmed/25988507 http://dx.doi.org/10.1038/srep10385 |
work_keys_str_mv | AT berclazcorinne combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT pachechristophe combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT bouwensarno combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT szlagdaniel combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT lopezantonio combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT joostenlieke combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT ekimselen combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT brommaarten combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT gotthardtmartin combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT grapinbottonanne combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers AT lassertheo combinedopticalcoherenceandfluorescencemicroscopytoassessdynamicsandspecificityofpancreaticbetacelltracers |