Shaping an evanescent focus of light for high spatial resolution optogenetic activations in live cells
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arXiv
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| Format: | Preprint |
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2024
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| author | Grosjean, Marc Grichine, Alexei Pezet, Mylene Destaing, Olivier Delon, Antoine Wang, Irène |
| author_facet | Grosjean, Marc Grichine, Alexei Pezet, Mylene Destaing, Olivier Delon, Antoine Wang, Irène |
| contents | Confining light illumination in the three dimensions of space is a challenge for various applications. Among these, optogenetic methods developed for live experiments in cell biology would benefit from such a localized illumination as it would improve the spatial resolution of diffusive photosensitive proteins leading to spatially constrained biological responses in specific subcellular organelles. Here, we describe a method to create and move a focused evanescent spot across the field of view of a high numerical aperture microscope objective, using a digital micro-mirror device (DMD). We show that, after correcting the optical aberrations, light is confined within a spot of sub-micron lateral size and $\sim$100~nm axial depth, resulting in a volume of illumination drastically smaller than the one generated by a standard propagative focus. This evanescent focus is sufficient to induce a more intense and localized recruitment compared to a propagative focus on the optogenetic system CRY2-CIBN, improving the resolution of its pattern of activation. |
| format | Preprint |
| id |
arxiv_https___arxiv_org_abs_2403_00699 |
| institution | arXiv |
| publishDate | 2024 |
| record_format | arxiv |
| spellingShingle | Shaping an evanescent focus of light for high spatial resolution optogenetic activations in live cells Grosjean, Marc Grichine, Alexei Pezet, Mylene Destaing, Olivier Delon, Antoine Wang, Irène Optics Biological Physics Confining light illumination in the three dimensions of space is a challenge for various applications. Among these, optogenetic methods developed for live experiments in cell biology would benefit from such a localized illumination as it would improve the spatial resolution of diffusive photosensitive proteins leading to spatially constrained biological responses in specific subcellular organelles. Here, we describe a method to create and move a focused evanescent spot across the field of view of a high numerical aperture microscope objective, using a digital micro-mirror device (DMD). We show that, after correcting the optical aberrations, light is confined within a spot of sub-micron lateral size and $\sim$100~nm axial depth, resulting in a volume of illumination drastically smaller than the one generated by a standard propagative focus. This evanescent focus is sufficient to induce a more intense and localized recruitment compared to a propagative focus on the optogenetic system CRY2-CIBN, improving the resolution of its pattern of activation. |
| title | Shaping an evanescent focus of light for high spatial resolution optogenetic activations in live cells |
| topic | Optics Biological Physics |
| url | https://arxiv.org/abs/2403.00699 |