Dynamic Ca²⁺ imaging with a simplified lattice light-sheet microscope: A sideways view of subcellular Ca²⁺ puffs.

ABSTRACT: We describe the construction of a simplified, inexpensive lattice light-sheet microscope, and illustrate its use for imaging subcellular Ca²⁺ puffs evoked by photoreleased i-IP₃ in cultured SH-SY5Y neuroblastoma cells loaded with the Ca²⁺ probe Cal520. The microscope provides sub-micron spatial resolution and enables recording of local Ca²⁺ transients in single-slice mode with a signal-to-noise ratio and temporal resolution (2ms) at least as good as confocal or total internal reflection microscopy. Signals arising from openings of individual IP₃R channels are clearly resolved, as are stepwise changes in fluorescence reflecting openings and closings of individual channels during puffs. Moreover, by stepping the specimen through the ligh

SUBMITTER: Ellefsen KL 

PROVIDER: S-EPMC5881921 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature