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Video 7: Rapid volumetric calcium imaging of highly motile Hydra.

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posted on 2018-05-10, 09:26 authored by Bianca Migliori, Malika S. Datta, Christophe Dupre, Mehmet C. Apak, Shoh Asano, Ruixuan Gao, Edward S. Boyden, Ola Hermanson, Rafael Yuste, Raju Tomer
GCaMP6s expressing Hydra was imaged using LSTM with 10x/0.6NA objective. Maximum intensity projections are shown for the two halves of the volume. First occurrences of longitudinal and radial contractions are annotated. The scale Bar is 100 microns.

An advantage of the LSTM imaging technique (see below for details) is the capacity for rapid volumetric live imaging of samples undergoing substantial non-isomorphic rearrangements in their body shape and cellular density, resulting in continuously changing local optical properties.

This video relates to the development of the imaging technique LSTM: Light Sheet Theta Microscopy for rapid high-resolution imaging of large biological samples, an approach designed to overcome the limitations of 'standard' Light Sheet microscopy LSM in terms of lateral dimensions and imaging quality, while preserving the benefits of LSM in terms of imaging resolution, depth and speed.

LSTM uses two symmetrically-arranged oblique light-sheets, generated using independent illumination objectives, for rapid high-resolution imaging of large samples.

View the full collection of related videos at:

https://doi.org/10.6084/m9.figshare.c.4072160

Funding

This work is mainly supported by Columbia University Arts & Sciences startup grant to RT. Hermanson445 Lab and BM are supported by VR-MH and BCF. RY and CD were supported by the NEI (DP1EY024503),446 and the reported Hydra material is based upon the work supported by the Defense Advanced Research447 Projects Agency (DARPA) under Contract No. HR0011-17-C-0026. In addition, Hydra research was448 supported in part by competitive fellowship funds from the H. Keffer Hartline, Edward F. MacNichol, Jr.449 Fellowship Fund, The E. E. Just Endowed Research Fellowship Fund, Lucy B. Lemann Fellowship Fund,450 and Frank R. Lillie Fellowship Fund Fellowship Fund of the Marine Biological Laboratory in Woods Hole,451 MA.

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Research Data Support

Research data support provided by Springer Nature.