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Shaping an evanescent focus of light for high spatial resolution optogenetic activations in live cells.
- Source :
-
Optics express [Opt Express] 2024 May 20; Vol. 32 (11), pp. 19480-19494. - Publication Year :
- 2024
-
Abstract
- 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, at the interface between a glass substrate and an aqueous sample, 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 ∼100 nm axial depth above the coverslip, 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.
- Subjects :
- Humans
Cryptochromes metabolism
Optogenetics methods
Light
Subjects
Details
- Language :
- English
- ISSN :
- 1094-4087
- Volume :
- 32
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Optics express
- Publication Type :
- Academic Journal
- Accession number :
- 38859082
- Full Text :
- https://doi.org/10.1364/OE.522639