title: Putting a finishing touch on GECIs.
abstract: More than a decade ago genetically encoded calcium indicators (GECIs) entered the stage as new promising tools to image calcium dynamics and neuronal activity in living tissues and designated cell types in vivo. From a variety of initial designs two have emerged as promising prototypes for further optimization: FRET (Förster Resonance Energy Transfer)-based sensors and single fluorophore sensors of the GCaMP family. Recent efforts in structural analysis, engineering and screening have broken important performance thresholds in the latest generation for both classes. While these improvements have made GECIs a powerful means to perform physiology in living animals, a number of other aspects of sensor function deserve attention. These aspects include indicator linearity, toxicity and slow response kinetics. Furthermore creating high performance sensors with optically more favorable emission in red or infrared wavelengths as well as new stably or conditionally GECI-expressing animal lines are on the wish list. When the remaining issues are solved, imaging of GECIs will finally have crossed the last milestone, evolving from an initial promise into a fully matured technology.
authors.affiliation: Max-Planck-Institute of Neurobiology Martinsried, Germany.
authors.forename: Tobias
authors.initials: T
authors.lastname: Rose
authors.affiliation: Max-Planck-Institute of Neurobiology Martinsried, Germany.
authors.forename: Pieter M
authors.initials: PM
authors.lastname: Goltstein
authors.affiliation: Max-Planck-Institute of Neurobiology Martinsried, Germany.
authors.forename: Ruben
authors.initials: R
authors.lastname: Portugues
authors.affiliation: Max-Planck-Institute of Neurobiology Martinsried, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Griesbeck
keywords: FRET; buffering; calcium; fluorescent protein; imaging; neuronal activity; segmentation