Abstract
Increasing the volumetric imaging speed of light-sheet microscopy will improve its ability to detect fast changes in neural activity. Here, a system is introduced for brain-wide imaging of neural activity in the larval zebrafish by coupling structured illumination with cubic phase extended depth-of-field (EDoF) pupil encoding. This microscope enables faster light-sheet imaging and facilitates arbitrary plane scanning-removing constraints on acquisition speed, alignment tolerances, and physical motion near the sample. The usefulness of this method is demonstrated by performing multi-plane calcium imaging in the fish brain with a 416×832×160  μm field of view at 33 Hz. The optomotor response behavior of the zebrafish is monitored at high speeds, and time-locked correlations of neuronal activity are resolved across its brain.
🔬 Techniques
🧬 Organisms
✨ Fluorophores
🧪 Sample Preparation
🏭 Microscope Brands
🧪 Reagent Suppliers
📷 Detectors
🔎 Objectives
💻 Software Details
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Fig. 1
Schematic of the excitation and imaging path is shown in panel A. The axial dependence for this realization of the cubic-phase point spread function is shown in panel B. The left-most panel is the sim...
Fig. 2
With conventional light-sheet microscopy, the tilt of the scanning beam results in out-of-focus imaging and loss of contrast of the scene in the focal plane. Panel A experimentally captures this loss ...
Fig. 3
Demonstration of functional imaging capabilities after data restoration. Panel A shows maximum-intensity projections conveying the spatial specificity of neurons responding to visual motion. Panel B s...
Fig. 4
Three axial planes are serially scanned in rapid succession to achieve 33 Hz sampling across a 160 u00b5m axial range. The restored planes are presented in Panel A. Temporal traces from individual cel...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
0 commentsNo comments yet. Be the first to start a discussion!
Leave a Comment