A Customized Light Sheet Microscope to Measure Spatio-Temporal Protein Dynamics in Small Model Organisms

Propagation of electromechanical waves in excitable heart muscles follows complex spatiotemporal patterns holding the key to understanding life-threatening arrhythmias and other cardiac conditions. Accurate volumetric mapping of cardiac wave propagation is currently hampered by fast heart motion, particularly in small model organisms. Here we demonstrate that ultrafast four-dimensional imaging of cardiac mechanical wave propagation in entire beating murine heart can be accomplished by sparse optoacoustic sensing with high contrast, ∼115-µm spatial and submillisecond temporal resolution. We extract accurate dispersion and phase velocity maps of the cardiac waves and reveal vortex-like patterns associated with mechanical phase singularities that occur during arrhythmic events induced via burst ventricular electric stimulation. The newly introduced cardiac mapping approach is a bold step toward deciphering the complex mechanisms underlying cardiac arrhythmias and enabling precise therapeutic interventions.

Download Full-text

Fully Automated Ultrasound-Based Touch Assay for Small Model Organisms

Biophysical Journal ◽

10.1016/j.bpj.2017.11.645 ◽

2018 ◽

Vol 114 (3) ◽

pp. 111a-112a

Author(s):

Miriam B. Goodman ◽

Holger Fehlauer

Keyword(s):

Model Organisms ◽

Small Model

Download Full-text

Light-sheet fluorescence imaging charts the gastrula origin of vascular endothelial cells in early zebrafish embryos

Cell Discovery ◽

10.1038/s41421-020-00204-7 ◽

2020 ◽

Vol 6 (1) ◽

Author(s):

Meijun Pang ◽

Linlu Bai ◽

Weijian Zong ◽

Xu Wang ◽

Ye Bu ◽

...

Keyword(s):

Endothelial Cells ◽

Fluorescence Imaging ◽

Vascular Endothelial Cells ◽

Cell Lineage ◽

Light Sheet ◽

Model Organisms ◽

Vascular Endothelial ◽

Anterior Trunk ◽

Potential Applications ◽

Complete Cell

Abstract It remains challenging to construct a complete cell lineage map of the origin of vascular endothelial cells in any vertebrate embryo. Here, we report the application of in toto light-sheet fluorescence imaging of embryos to trace the origin of vascular endothelial cells (ECs) at single-cell resolution in zebrafish. We first adapted a previously reported method to embryo mounting and light-sheet imaging, created an alignment, fusion, and extraction all-in-one software (AFEIO) for processing big data, and performed quantitative analysis of cell lineage relationships using commercially available Imaris software. Our data revealed that vascular ECs originated from broad regions of the gastrula along the dorsal–ventral and anterior–posterior axes, of which the dorsal–anterior cells contributed to cerebral ECs, the dorsal–lateral cells to anterior trunk ECs, and the ventral–lateral cells to posterior trunk and tail ECs. Therefore, this work, to our knowledge, charts the first comprehensive map of the gastrula origin of vascular ECs in zebrafish, and has potential applications for studying the origin of any embryonic organs in zebrafish and other model organisms.

Download Full-text

Automated spatio-temporal analysis of dendritic spines and related protein dynamics

PLoS ONE ◽

10.1371/journal.pone.0182958 ◽

2017 ◽

Vol 12 (8) ◽

pp. e0182958 ◽

Cited By ~ 3

Author(s):

Vincent On ◽

Atena Zahedi ◽

Iryna M. Ethell ◽

Bir Bhanu

Keyword(s):

Protein Dynamics ◽

Dendritic Spines ◽

Temporal Analysis ◽

Related Protein ◽

Spatio Temporal

Download Full-text

Applying dimorphic yeasts as model organisms to study mycelial growth: Part 1. Experimental investigation of the spatio-temporal development of filamentous yeast colonies

Bioprocess and Biosystems Engineering ◽

10.1007/s00449-010-0442-6 ◽

2010 ◽

Vol 34 (1) ◽

pp. 13-20 ◽

Cited By ~ 4

Author(s):

Thomas Walther ◽

Holger Reinsch ◽

Philipp Weber ◽

Kai Ostermann ◽

Andreas Deutsch ◽

...

Keyword(s):

Experimental Investigation ◽

Mycelial Growth ◽

Model Organisms ◽

Temporal Development ◽

Spatio Temporal

Download Full-text

High-Resolution, Large Field-of-View, and Multi-View Single Objective Light-Sheet Microscopy

10.1101/2020.09.22.309229 ◽

2020 ◽

Author(s):

Bin Yang ◽

Alfred Millett-Sikking ◽

Merlin Lange ◽

Ahmet Can Solak ◽

Hirofumi Kobayashi ◽

...

Keyword(s):

High Resolution ◽

Light Sheet ◽

Field Of View ◽

Large Field ◽

Sample Rotation ◽

Light Sheet Microscopy ◽

Spatio Temporal ◽

Large Living ◽

Single Objective ◽

Imaging Speed

Light-sheet microscopy has become the preferred method for long-term imaging of large living samples because of its low photo-invasiveness and good optical sectioning capabilities. Unfortunately, refraction and scattering often pose obstacles to light-sheet propagation and limit imaging depth. This is typically addressed by imaging multiple complementary views to obtain high and uniform image quality throughout the sample. However, multi-view imaging often requires complex multi-objective configurations that complicate sample mounting, or sample rotation that decreases imaging speed. Recent developments in single-objective light-sheet microscopy have shown that it is possible to achieve high spatio-temporal resolution with a single objective for both illumination and detection. Here we describe a single-objective light-sheet microscope that achieves: (i) high-resolution and large field-of-view imaging via a custom remote focusing objective; (ii) simpler design and ergonomics by remote placement of coverslips; (iii) fast volumetric imaging by means of light-sheet stabilised stage scanning – a novel scanning modality that extends the imaging volume without compromising imaging speed nor quality; (iv) multi-view imaging by means of dual orthogonal light-sheet illumination. Finally, we demonstrate the speed, field of view and resolution of our novel instrument by imaging zebrafish tail development.

Download Full-text

Light-Sheet Fluorescence Imaging Charts the Gastrula Origin of Vascular Endothelial Cells in Early Zebrafish Embryos

10.1101/2020.05.27.118661 ◽

2020 ◽

Author(s):

Meijun Pang ◽

Linlu Bai ◽

Weijian Zong ◽

Xu Wang ◽

Ye Bu ◽

...

Keyword(s):

Endothelial Cells ◽

Fluorescence Imaging ◽

Vascular Endothelial Cells ◽

Cell Lineage ◽

Light Sheet ◽

Model Organisms ◽

Vascular Endothelial ◽

Anterior Trunk ◽

Potential Applications ◽

Complete Cell

AbstractIt remains challenging to construct a complete cell lineage map of the origin of vascular endothelial cells in any vertebrate embryo. Here, we report the application of in toto light-sheet fluorescence imaging of embryos to tracing the origin of vascular endothelial cells (ECs) at single-cell resolution in zebrafish. We first adapted a previously-reported method to mount embryos and light-sheet imaging, created an alignment, fusion, and extraction all-in-one software (AFEIO) for processing big data, and performed quantitative analysis of cell lineage relationships using commercially-available Imaris software. Our data revealed that vascular ECs originated from broad regions of the gastrula along the dorsal-ventral and anterior-posterior axes, of which the dorsal-anterior cells contributed to cerebral ECs, the dorsal-lateral cells to anterior trunk ECs, and the ventral-lateral cells to posterior trunk and tail ECs. Therefore, this work, to our knowledge, charts the first comprehensive map of the gastrula origin of vascular ECs in zebrafish, and has potential applications for studying the origin of any embryonic organs in zebrafish and other model organisms.

Download Full-text

3D Interferometric Lattice Light-Sheet Imaging

10.1101/2020.08.27.266999 ◽

2020 ◽

Author(s):

Bin Cao ◽

Guanshi Wang ◽

Jieru Li ◽

Alexandros Pertsinidis

Keyword(s):

Single Molecule ◽

Super Resolution ◽

Light Sheet ◽

Detection Sensitivity ◽

Live Cells ◽

Background Suppression ◽

Axial Resolution ◽

Volumetric Imaging ◽

Spatio Temporal ◽

And Function

Understanding cellular structure and function requires live-cell imaging with high spatio-temporal resolution and high detection sensitivity. Direct visualization of molecular processes using single-molecule/super-resolution techniques has thus been transformative. However, extracting the highest-resolution 4D information possible from weak and dynamic fluorescence signals in live cells remains challenging. For example, some of the highest spatial resolution methods, e.g. interferometric (4Pi) approaches1-6 can be slow, require high peak excitation intensities that accelerate photobleaching or suffer from increased out-of-focus background. Selective-plane illumination (SPIM)7-12 reduces background, but most implementations typically feature modest spatial, especially axial, resolution. Here we develop 3D interferometric lattice light-sheet (3D-iLLS) imaging, a technique that overcomes many of these limitations. 3D-iLLS provides, by virtue of SPIM, low light levels and photobleaching, while providing increased background suppression and significantly improved volumetric imaging/sectioning capabilities through 4Pi interferometry. We demonstrate 3D-iLLS with axial resolution and single-particle localization precision down to <100nm (FWHM) and <10nm (1σ) respectively. 3D-iLLS paves the way for a fuller elucidation of sub-cellular phenomena by enhanced 4D resolution and SNR live imaging.

Download Full-text