Imaging neural activity in zebrafish larvae with adaptive optics and structured illumination light sheet microscopy

2019 ◽  
Author(s):  
Peter Kner ◽  
Yang Liu ◽  
Aqsa Malik ◽  
Keelan Lawrence ◽  
Chelsea E. Gunderson ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Neural Activity ◽  
Light Sheet ◽  
Structured Illumination ◽  
Light Sheet Microscopy ◽  
Zebrafish Larvae

scholarly journals Dual-view light-sheet imaging through tilted glass interface using a deformable mirror

2020 ◽  
Author(s):  
N Vladimirov ◽  
F Preusser ◽  
J Wisniewski ◽  
Z Yaniv ◽  
RA Desai ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
High Speed ◽  
Microfluidic Channel ◽  
Microfluidic Devices ◽  
Light Sheet ◽  
Stochastic Gradient Descent ◽  
Deformable Mirror ◽  
Optical Aberrations ◽  
Light Sheet Microscopy ◽  
Gradient Descent Algorithm

AbstractLight-sheet microscopy has become one of the primary tools for imaging live developing organisms because of its high speed, low phototoxicity, and optical sectioning capabilities. Detection from multiple sides (multi-view imaging) additionally allows nearly isotropic resolution via computational merging of the views. However, conventional light-sheet microscopes require that the sample is suspended in a gel to allow optical access from two or more sides. At the same time, the use of microfluidic devices is highly desirable for many experiments, but geometric constrains and strong optical aberrations caused by the coverslip titled relative to objectives make the use of multi-view lightsheet challenging for microfluidics.In this paper we describe the use of adaptive optics (AO) to enable multi-view light-sheet microscopy in such microfluidic setup by correcting optical aberrations introduced by the tilted coverslip. The optimal shape of deformable mirror is computed by an iterative stochastic gradient-descent algorithm that optimizes PSF in two orthogonal planes simultaneously. Simultaneous AO correction in two optical arms is achieved via a knife-edge mirror that splits excitation path and combines the detection path.We characterize the performance of this novel microscope setup and, by dual-view light-sheet imaging of C.elegans inside a microfluidic channel, demonstrate a drastic improvement of image quality due to AO and dual-view reconstruction. Our microscope design allows multi-view light-sheet microscopy with microfluidic devices for precisely controlled experimental conditions and high-content screening.

scholarly journals A polymer gel index-matched to water enables diverse applications in fluorescence microscopy

2020 ◽  
Author(s):  
Xiaofei Han ◽  
Yijun Su ◽  
Hamilton White ◽  
Kate M. O’Neill ◽  
Nicole Y. Morgan ◽  
...  
Keyword(s):  
Surface Passivation ◽  
Super Resolution ◽  
Light Sheet ◽  
Polymer Gel ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Neural Structure ◽  
Uv Curable ◽  
Isotropic Resolution ◽  
Light Sheet Microscopy

AbstractWe demonstrate diffraction-limited and super-resolution imaging through thick layers (tens-hundreds of microns) of BIO-133, a biocompatible, UV-curable, commercially available polymer with a refractive index (RI) matched to water. We show that cells can be directly grown on BIO-133 substrates without the need for surface passivation and use this capability to perform extended time-lapse volumetric imaging of cellular dynamics 1) at isotropic resolution using dual-view light-sheet microscopy, and 2) at super-resolution using instant structured illumination microscopy. BIO-133 also enables immobilization of 1) Drosophila tissue, allowing us to track membrane puncta in pioneer neurons, and 2) Caenorhabditis elegans, which allows us to image and inspect fine neural structure and to track pan-neuronal calcium activity over hundreds of volumes. Finally, BIO-133 is compatible with other microfluidic materials, enabling optical and chemical perturbation of immobilized samples, as we demonstrate by performing drug and optogenetic stimulation on cells and C. elegans.

scholarly journals Observing the Cell in Its Native State: Imaging Subcellular Dynamics in Multicellular Organisms

2018 ◽  
Author(s):  
Tsung-Li Liu ◽  
Srigokul Upadhyayula ◽  
Daniel E. Milkie ◽  
Ved Singh ◽  
Kai Wang ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
High Speed ◽  
Developmental Stages ◽  
Phenotypic Diversity ◽  
Metastatic Cancer ◽  
Light Sheet ◽  
Intrinsic Variability ◽  
Spatiotemporal Resolution ◽  
Light Sheet Microscopy

AbstractTrue physiological imaging of subcellular dynamics requires studying cells within their parent organisms, where all the environmental cues that drive gene expression, and hence the phenotypes we actually observe, are present. A complete understanding also requires volumetric imaging of the cell and its surroundings at high spatiotemporal resolution without inducing undue stress on either. We combined lattice light sheet microscopy with two-channel adaptive optics to achieve, across large multicellular volumes, noninvasive aberration-free imaging of subcellular processes, including endocytosis, organelle remodeling during mitosis, and the migration of axons, immune cells, and metastatic cancer cells in vivo. The technology reveals the phenotypic diversity within cells across different organisms and developmental stages, and may offer insights into how cells harness their intrinsic variability to adapt to different physiological environments.One Sentence SummaryCombining lattice light sheet microscopy with adaptive optics enables high speed, high resolution in vivo 3D imaging of dynamic processes inside cells under physiological conditions within their parent organisms.

Adaptive optics light-sheet microscopy for functional neuroimaging

2021 ◽  
Author(s):  
Antoine Hubert ◽  
Fabrice Harms ◽  
Sophie Imperato ◽  
Vincent Loriette ◽  
Cynthia Veilly ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Functional Neuroimaging ◽  
Light Sheet ◽  
Light Sheet Microscopy

scholarly journals Lamellar junctions in the endolymphatic sac act as a relief valve to regulate inner ear pressure

2017 ◽  
Author(s):  
Ian A. Swinburne ◽  
Kishore R. Mosaliganti ◽  
Srigokul Upadhyayula ◽  
Tsung-Li Liu ◽  
David G. C. Hildebrand ◽  
...  
Keyword(s):  
Inner Ear ◽  
Adaptive Optics ◽  
Ion Homeostasis ◽  
Fluid Pressure ◽  
Light Sheet ◽  
Normal Function ◽  
Endolymphatic Sac ◽  
Fluid Composition ◽  
Relief Valve ◽  
Light Sheet Microscopy

AbstractThe inner ear is a fluid-filled closed-epithelial structure whose normal function requires maintenance of an internal hydrostatic pressure and fluid composition by unknown mechanisms. The endolymphatic sac (ES) is a dead-end epithelial tube connected to the inner ear. ES defects can cause distended ear tissue, a pathology often seen in hearing and balance disorders. Using live imaging of zebrafish larvae, we reveal that the ES undergoes cycles of slow pressure-driven inflation followed by rapid deflation every 1-3 hours. Using serial-section electron microscopy and adaptive optics lattice light-sheet microscopy, we find a pressure relief valve in the ES comprised of thin overlapping basal lamellae that dynamically extend over neighboring cells before rupturing under pressure leading to ES collapse. The unexpected discovery of a physical relief valve in the ear emphasizes the need for further study into how organs control fluid pressure, volume, flow, and ion homeostasis in development and disease.

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