JoVE Journal

Developmental Biology

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Isotropic Light-Sheet Microscopy and Automated Cell Lineage Analyses to Catalogue Caenorhabditis elegans Embryogenesis with Subcellular Resolution

8.1K Views

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08:16 min

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June 6th, 2019

DOI :

10.3791/59533-v

June 6th, 2019

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Leighton H. Duncan¹^,⁵, Mark W. Moyle¹^,⁵, Lin Shao¹^,⁵, Titas Sengupta¹^,⁵, Richard Ikegami¹^,⁵, Abhishek Kumar⁴^,⁵, Min Guo⁴^,⁵, Ryan Christensen⁴^,⁵, Anthony Santella²^,⁵, Zhirong Bao²^,⁵, Hari Shroff⁴^,⁵, William Mohler³^,⁵, Daniel A. Colón-Ramos¹^,⁵^,⁶

¹Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine, ²Developmental Biology Program, Sloan Kettering Institute, ³Department of Genetics and Genome Sciences and Center for Cell Analysis and Modeling, University of Connecticut Health Center, ⁴Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, ⁵WormGUIDES.org, ⁶Instituto de Neurobiología, Recinto de Ciencias Médicas, Universidad de Puerto Rico

Summary

Here, we present a combinatorial approach using high-resolution microscopy, computational tools, and single-cell labeling in living C. elegans embryos to understand single cell dynamics during neurodevelopment.

Explore More Videos

Isotropic Light Sheet Microscopy

Automated Cell Lineage Analysis

Caenorhabditis Elegans

Dual view Inverted Selective Plane Illumination Microscopy

Signal To Noise Ratio

Subcellular Resolution

Cell Morphology

Microcapillary Pipette

Imaging Chamber

Laser Intensity Calibration

Autofocus Analysis

Nuclear Fluorescence Channel

Chapters in this video

0:04

Title

0:34

C. elegans Embryo Sample Mounting Preparation

1:48

Dual-View Inverted Selective Plane Illumination Microscopy (diSPIM) Embryonic Imaging

3:12

Isotropic High-Resolution Volumetric Data: Cell Lineage Analysis

6:06

Results: Representative Cellular Identification and Single Cell Characterization of Neurite Outgrowth Dynamics

7:43

Conclusion

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