The development of fluorescent indicators capable of reporting intracellular Ca2+ ion concentration [17] allowed a new series of RF exposure experiments to be performed using fluorescence microspectroscopy methods.
Alterations in the polymethine chain allow the construction of fluorescent indicators.
It consists in a specific design of fluorescent indicator using Excited-State Intramolecular Proton-Transfer (ESIPT) phenomenon and enhancement of its fluorescence on the metal surface (Metal Enhanced Fluorescence – MEF).
Due to the breath of literature citing intracellular Ca2+ as a secondary messenger, the use of fluorescent calcium indicators is chosen as the means of observing endothelial sensory function.
Direct detection of reactive oxygen species (ROS) involved in cellular oxidative stress in living cells has relied heavily on the use of the fluorescent indicator 2,7-dichlorodihydrofluorescein diacetate (H2DCF-DA) (Crow 1997).
Traditional methods of assessing intracellular Ca2+ handling, such as use of Ca2+-sensitive fluorescent indicators and confocal microscopy, can be applied to iPSC-CMs to confirm the presence of intact and functional Ca2+ handling apparatus (figure 3D).
Nowadays, techniques that use fluorescent indicators allow us to measure the cytosolic Ca2+ concentration in several models of smooth muscles [ 50].
Proton transfer is an important player in this respect and is central to the use of fluorescent proteins (FPs) as pH indicators and as reversibly photoswitchable dyes utilized in super-resolution microscopy.
