Bioluminescence imaging of calcium oscillations inside intracellular organelles.
Ca(2+) oscillations inside intracellular organelles are important for regulation of functions such as gene expression at the nucleus, respiration at mitochondria or protein processing at the endoplasmic reticulum. Targeted aequorins are excellent calcium probes for subcellular analysis, but single-cell imaging has proven difficult because of low light yield. Here we describe a procedure that combines virus-based expression of targeted aequorins with photon-counting imaging. This methodology allows real-time resolution of changes of cytosolic, mitochondrial or nuclear Ca(2+) signals at the single-cell level.
Villalobos
Carlos
C
Instituto de Biología y Genética Molecular, Universidad de Valladolid and Consejo Superior de Investigaciones Científicas, Valladolid, Spain.
Alonso
María Teresa
MT
García-Sancho
Javier
J
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D007515:Islets of Langerhans / Q000378:metabolism; D049449:Luminescence; D051379:Mice; D008807:Mice, Inbred BALB C; D015388:Organelles / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
D002118:Calcium