The internal brain dynamics that link sensation and action are arguably better studied during natural animal behaviors. Here, we report on a novel volume imaging and 3D tracking technique that monitors whole brain neural activity in freely swimming larval zebrafish (Danio rerio). We demonstrated the capability of our system through functional imaging of neural activity during visually evoked and prey capture behaviors in larval zebrafish.
23The internal brain dynamics that link sensation and action are arguably better studied 24 during natural animal behaviors. Here we report on a novel volume imaging and 3D 25 tracking technique that monitors whole brain neural activity in freely swimming larval 26 zebrafish (Danio rerio). We demonstrated the capability of our system through functional 27 imaging of neural activity during visually evoked and prey capture behaviors in larval 28 zebrafish. 29 30 31
One of the major challenges in large scale optical imaging of neuronal
activity is to simultaneously achieve sufficient temporal and spatial
resolution across a large volume. Here, we introduce sparse
decomposition light-field microscopy (SDLFM), a computational imaging
technique based on light-field microscopy (LFM) that takes algorithmic
advantage of the high temporal resolution of LFM and the inherent
temporal sparsity of spikes to improve effective spatial resolution
and signal-to-noise ratios (SNRs). With increased effective spatial
resolution and SNRs, neuronal activity at the single-cell level can be
recovered over a large volume. We demonstrate the single-cell imaging
capability of SDLFM with in vivo imaging
of neuronal activity of whole brains of larval zebrafish with
estimated lateral and axial resolutions of
∼
3.5
µ
m
and
∼
7.4
µ
m
, respectively, acquired at volumetric
imaging rates up to 50 Hz. We also show that SDLFM increases the
quality of neural imaging in adult fruit flies.
Monitoring and manipulating neuronal activities with optical microscopy desires a method where light can be focused or projected over a long axial range so that large brain tissues (>100 [Formula: see text] thick) can be simultaneously imaged, and specific brain regions can be optogenetically stimulated without the need for slow optical refocusing. However, the micron-scale resolution required in neuronal imaging yields a depth of field of less than 10 [Formula: see text] in conventional imaging systems. We propose to use a circularly symmetric phase mask to extend the depth of field. A numerical study shows that our method maintains both the peak and the shape of the point spread function vs the axial position better than current methods. Imaging of a 3D bead suspension and sparsely labelled thick brain tissue confirms the feasibility of the system for fast volumetric imaging.
Molecular signals interact to mediate diverse biological computations. Ideally one would be able to image many signals at once, in the same living cell, to reveal how they work together. Here we report temporally multiplexed imaging (TMI), which uses the clocklike properties of fluorescent proteins to enable different cellular signals to be represented by different temporal fluorescence codes. Using different photoswitchable fluorescent proteins to represent different cellular signals, we can linearly decompose a brief movie of the fluorescence fluctuations in a given cell, into a sum of the fluctuation traces of each individual fluorophore, each weighted by its respective signal amplitude. We demonstrate the power of TMI to report relationships amongst a diversity of second messenger, kinase, and cell cycle signals, using ordinary microscopes.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.