Abstract
Cells in the brain act as components of extended networks. Therefore, to understand neurobiological processes in a physiological context, it is essential to study them in vivo. Super-resolution microscopy has spatial resolution beyond the diffraction limit, thus promising to provide structural and functional insights that are not accessible with conventional microscopy. However, to apply it to in vivo brain imaging, we must address the challenges of 3D imaging in an optically heterogeneous tissue that is constantly in motion. We optimized image acquisition and reconstruction to combat sample motion and applied adaptive optics to correcting sample-induced optical aberrations in super-resolution structured illumination microscopy (SIM) in vivo. We imaged the brains of live zebrafish larvae and mice and observed the dynamics of dendrites and dendritic spines at nanoscale resolution.
Highlights
Cells in the brain act as components of extended networks
The optical system for in vivo structured illumination microscopy (SIM) imaging of the brain consisted of two modules: one for SIM itself and one for adaptive optics (AO) to compensate for specimen-induced aberrations
The light passed through the AO module, where it reflected off a pupil-conjugate deformable mirror before entering the objective lens pupil to excite patterned fluorescence over a wide field within the sample (SI Appendix, Fig. S1C)
Summary
Cells in the brain act as components of extended networks. to understand neurobiological processes in a physiological context, it is essential to study them in vivo. We combined adaptive optics (AO) with SIM to eliminate sample-induced aberrations, used subdiffractive objects to accurately evaluate illumination parameters, and used phase up-sampling and image registration to alleviate the effect of brain motion.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
