Journal: STAR Protocols

Article Title: Two-photon manipulation of neuronal activity and behavior in Hydra vulgaris

doi: 10.1016/j.xpro.2023.102453

Figure Lengend Snippet:

Article Snippet: Note: Example microscope setup (used in Yamamoto and Yuste, 2023) – A modified two-photon laser scanning microscope (Ultima In Vivo, Bruker) was used.

Techniques: Transgenic Assay, Recombinant, Software, Microscopy, Imaging, Laser-Scanning Microscopy, In Vivo

Journal: bioRxiv

Article Title: The Reissner fiber under tension in vivo shows dynamic interaction with ciliated cells contacting the cerebrospinal fluid

doi: 10.1101/2023.02.22.529498

Figure Lengend Snippet: (A1) Time-series standard deviation projection in the sagittal plane from 2 photon laser scanning microscope showing the signal from the Reissner fiber and CSF-cNs in the central canal of 3 dpf Tg ( sspo:sspo-GFP;pkd2l1:tagRFP;pkd2l1:GCaMP5G ) zebrafish larva before RF photoablation. (A2) Time-series standard deviation projection in the sagittal plane from 2 photon laser scanning microscope showing the signal from CSF-cNs after RF ablation performed by spiral scanning photoablation with an infrared pulsed laser tuned at 800 nm over 0.5 μm on the RF (see Methods). (B) ROI selection for ventral CSF-cNs to analyze activity before and after RF photoablation within the same cells (top). Example calcium activity traces normalized to baseline for each of the ROIs before (red) and after (black) RF photoablation over 75 s imaged at 3.45 Hz (see Methods). (C) Percentage of active ventral CSF-cNs (active is defined as having at least 1 calcium transient for the recording) before and after RF photoablation (109 cells total from 8 fish from 2 independent clutches; mean percent active before ablation = 27.72% ± 6.34% versus mean percent active after ablation = 10.59% ± 3.1%; paired two-tailed t-test: p < 0.05). (D) The ratio of active ventral CSF-cNs after RF photoablation to those active before RF photoablation, illustrating on average, a fraction (on average ± SEM: 45% ± 14%) of active ventral CSF-cNs before photoablation remain active after RF photoablation. The purple lines on the graph represent the mean and the error bars indicate the SEM. (E) Pie charts illustrating the number of events per ventral CSF-cN before and after RF photoablation (mean number of events in active cells before RF photoablation = 0.94 events / min versus mean number of events in active cells after RF photoablation = 0.87 events / min; paired two-tailed t-test: p < 0.005). *p < 0.05, ** p < 0.005 Scale bar is 20 μm (A1, A2), 10 μm (B).

Article Snippet: Two-photon spiral scanning ablations at 800 nm were performed with a two-photon laser scanning microscope (2p- vivo , Intelligent Imaging Innovations, Inc., Denver, Colorado, USA) equipped with a 20X objective (N.A.

Techniques: Standard Deviation, Laser-Scanning Microscopy, Selection, Activity Assay, Two Tailed Test