Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment
The majority of intravital studies on brain tumor in living animal so far rely on dual color imaging. We describe here a multiphoton imaging protocol to dynamically characterize the interactions between six cellular components in a living mouse. We applied this methodology to a clinically relevant g...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2014-02-01
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| Series: | Frontiers in Cellular Neuroscience |
| Subjects: | |
| Online Access: | http://journal.frontiersin.org/Journal/10.3389/fncel.2014.00057/full |
| _version_ | 1828350938981597184 |
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| author | Clément eRicard Clément eRicard Clément eRicard Franck Christian Debarbieux Franck Christian Debarbieux Franck Christian Debarbieux |
| author_facet | Clément eRicard Clément eRicard Clément eRicard Franck Christian Debarbieux Franck Christian Debarbieux Franck Christian Debarbieux |
| author_sort | Clément eRicard |
| collection | DOAJ |
| description | The majority of intravital studies on brain tumor in living animal so far rely on dual color imaging. We describe here a multiphoton imaging protocol to dynamically characterize the interactions between six cellular components in a living mouse. We applied this methodology to a clinically relevant glioblastoma multiforme (GBM) model designed in reporter mice with targeted cell populations labeled by fluorescent proteins of different colors. This model permitted us to make non-invasive longitudinal and multi-scale observations of cell-to-cell interactions. We provide examples of such 5D (x,y,z,t,color) images acquired on a daily basis from volumes of interest, covering most of the mouse parietal cortex at subcellular resolution. Spectral deconvolution allowed us to accurately separate of each cell population as well as some components of the extracellular matrix. The technique represents a powerful tool for investigating how tumor progression is influenced by the interactions of tumor cells with host cells and the extracellular matrix micro-environment. It will be especially valuable for evaluating neuro-oncological drug efficacy and target specificity. The imaging protocol provided here can be easily translated to other mouse models of neuropathologies, and should also be of fundamental interest for investigations in other areas of systems biology. |
| first_indexed | 2024-04-14T01:32:50Z |
| format | Article |
| id | doaj.art-97912d9f03464292843f2fab0453df4f |
| institution | Directory Open Access Journal |
| issn | 1662-5102 |
| language | English |
| last_indexed | 2024-04-14T01:32:50Z |
| publishDate | 2014-02-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular Neuroscience |
| spelling | doaj.art-97912d9f03464292843f2fab0453df4f2022-12-22T02:20:06ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022014-02-01810.3389/fncel.2014.0005772866Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironmentClément eRicard0Clément eRicard1Clément eRicard2Franck Christian Debarbieux3Franck Christian Debarbieux4Franck Christian Debarbieux5Aix-Marseille University - CNRSAix-Marseille UniversityAix-Marseille University - CNRSAix-Marseille University - CNRSAix-Marseille UniversityAix-Marseille University - CNRSThe majority of intravital studies on brain tumor in living animal so far rely on dual color imaging. We describe here a multiphoton imaging protocol to dynamically characterize the interactions between six cellular components in a living mouse. We applied this methodology to a clinically relevant glioblastoma multiforme (GBM) model designed in reporter mice with targeted cell populations labeled by fluorescent proteins of different colors. This model permitted us to make non-invasive longitudinal and multi-scale observations of cell-to-cell interactions. We provide examples of such 5D (x,y,z,t,color) images acquired on a daily basis from volumes of interest, covering most of the mouse parietal cortex at subcellular resolution. Spectral deconvolution allowed us to accurately separate of each cell population as well as some components of the extracellular matrix. The technique represents a powerful tool for investigating how tumor progression is influenced by the interactions of tumor cells with host cells and the extracellular matrix micro-environment. It will be especially valuable for evaluating neuro-oncological drug efficacy and target specificity. The imaging protocol provided here can be easily translated to other mouse models of neuropathologies, and should also be of fundamental interest for investigations in other areas of systems biology.http://journal.frontiersin.org/Journal/10.3389/fncel.2014.00057/fullbrain tumorstwo-photon microscopycellular interactionscranial windowintravital imagingmulticolor transgenic mouse |
| spellingShingle | Clément eRicard Clément eRicard Clément eRicard Franck Christian Debarbieux Franck Christian Debarbieux Franck Christian Debarbieux Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment Frontiers in Cellular Neuroscience brain tumors two-photon microscopy cellular interactions cranial window intravital imaging multicolor transgenic mouse |
| title | Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment |
| title_full | Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment |
| title_fullStr | Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment |
| title_full_unstemmed | Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment |
| title_short | Six-color intravital two-photon imaging of brain tumors and their dynamic microenvironment |
| title_sort | six color intravital two photon imaging of brain tumors and their dynamic microenvironment |
| topic | brain tumors two-photon microscopy cellular interactions cranial window intravital imaging multicolor transgenic mouse |
| url | http://journal.frontiersin.org/Journal/10.3389/fncel.2014.00057/full |
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