Lightfield 4D Roadshow

Cleared spheroid of a co-culture of HCT-116-GFP (colon cancer) / NIH-3T3-RFP (fibroblasts) cells stained with Hoechst for nuclei. Imaged in an InSphero Akura plate. Dataset was segmented using arivis Pro. Sample courtesy of InSphero AG. Schlieren, Switzerland

Imaging calcium signaling as proxy for neuron activity is a widely used technique in many model systems. These signals occur rapidly, in milliseconds, requiring high temporal resolution.
The video shows calcium signaling in the zebrafish brain. Thanks to the large volume and speed of Lightfield 4D, neurons more than 50 µm apart from each other can be recorded at the same time. Additional high-resolution data was acquired using the Airyscan CO-8Y mode.
Data recorded from a zebrafish larvae 4 days post fertilization expressing the calcium reporter GCaMP6; image volume: 361 × 361 × 109 µm³; 10 volumes per second for 1 minute (661 time points); exposure time 91 ms; intensity coding LUT (low intensity blue, high intensity red to white).
Courtesy of Anton Nikolaev, University of Sheffield, UK. Data acquired at Wolfson Light Microscopy Facility in the School of Biosciences at the University of Sheffield.