Abstract

Following Ca²⁺ Changes in Acutely Stimulated Spheroids

Fluorescent confocal and light-sheet live Ca²⁺ imaging

Spheroids are easy and cost-effective 3D cell cultures, useful for fundamental and applied research. They can bridge the gap between 2D and in vivo experiments and can be used for high-throughput screenings. However, performing live microscopy to visualize acute drug responses is difficult with these roundish and non-adherent cell aggregates, hampering the study of rapid changes in intra- and intercellular signalling. To solve this issue, we stabilized spheroids in micro-perfusion chambers using a transparent and soft gelatine mesh.

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This approach allowed us to image Ca2+ changes by live confocal microscopy using an inverted TCS SP8 confocal microscope upon acute perfusion. Spheroids of immortalized cells from human taste/tongue papillae (BRAIN Biotech AG) were stimulated with tastants or ATP. Analysing the responses in concentric spheroid regions showed that cells at the rim displayed larger and faster transients than those in the core. These regional differences were confirmed in analogous experiments with light-sheet microscopy, done with a TCS SP8 DLS, which allowed us to image approximately 100 μm in depth without losing spatio-temporal resolution.

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Molitor E., Nürnberg E., Ertongur-Fauth T., Scholz P., Riedel K., Hafner M., Rudolf R. & Cesetti T.:

Analysis of Calcium Signaling in Live Human Tongue Cell 3D-Cultures upon Tastant Perfusion

Cell Calcium 87 (May 2020): 102164, DOI: 10.1016/j.ceca.2020.102164. https://www.sciencedirect.com/science/article/abs/pii/S0143416020300063

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