Imaging protein interactions in vivo with sub-cellular resolution
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| Main Authors: | , , , , , , , , |
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| Format: | Preprint |
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2008
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| _version_ | 1866917828181884928 |
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| author | Raicu, Valerica Stoneman, Michael R. Fung, Russell Melnichuk, Mike Jansma, David B. Pisterzi, Luca Fox, Michael Wells, James W. Saldin, Dilano K. |
| author_facet | Raicu, Valerica Stoneman, Michael R. Fung, Russell Melnichuk, Mike Jansma, David B. Pisterzi, Luca Fox, Michael Wells, James W. Saldin, Dilano K. |
| contents | Resonant Energy Transfer (RET) from an optically excited donor molecule (D) to a non-excited acceptor molecule (A) residing nearby is widely used to detect molecular interactions in living cells. Stoichiometric information, such as the number of proteins forming a complex, has been obtained so far for a handful of proteins, but only after exposing the sample sequentially to at least two different excitation wavelengths. During this lengthy process of measurement, the molecular makeup of a cellular region may change, and this has so far limited the applicability of RET to determination of cellular averages. Here we demonstrate a method for imaging protein complex distribution in living cells with sub-cellular spatial resolution, which relies on a spectrally-resolved two-photon microscope, a simple but competent theory, and a keen selection of fluorescent tags. This technology may eventually lead to tracking dynamics of macromolecular complex formation and dissociation with spatial resolution inside living cells. |
| format | Preprint |
| id |
arxiv_https___arxiv_org_abs_0810_2341 |
| institution | arXiv |
| publishDate | 2008 |
| record_format | arxiv |
| spellingShingle | Imaging protein interactions in vivo with sub-cellular resolution Raicu, Valerica Stoneman, Michael R. Fung, Russell Melnichuk, Mike Jansma, David B. Pisterzi, Luca Fox, Michael Wells, James W. Saldin, Dilano K. Biological Physics Optics Resonant Energy Transfer (RET) from an optically excited donor molecule (D) to a non-excited acceptor molecule (A) residing nearby is widely used to detect molecular interactions in living cells. Stoichiometric information, such as the number of proteins forming a complex, has been obtained so far for a handful of proteins, but only after exposing the sample sequentially to at least two different excitation wavelengths. During this lengthy process of measurement, the molecular makeup of a cellular region may change, and this has so far limited the applicability of RET to determination of cellular averages. Here we demonstrate a method for imaging protein complex distribution in living cells with sub-cellular spatial resolution, which relies on a spectrally-resolved two-photon microscope, a simple but competent theory, and a keen selection of fluorescent tags. This technology may eventually lead to tracking dynamics of macromolecular complex formation and dissociation with spatial resolution inside living cells. |
| title | Imaging protein interactions in vivo with sub-cellular resolution |
| topic | Biological Physics Optics |
| url | https://arxiv.org/abs/0810.2341 |