se0c00277_si_001.pdf (1.88 MB)
Assessment of a Silicon-Photomultiplier-Based Platform for the Measurement of Intracellular Calcium Dynamics with Targeted Aequorin
journal contribution
posted on 2020-08-05, 19:04 authored by Federico
Alessandro Ruffinatti, Samuela Lomazzi, Luca Nardo, Romualdo Santoro, Alexander Martemiyanov, Marianna Dionisi, Laura Tapella, Armando A. Genazzani, Dmitry Lim, Carla Distasi, Massimo CacciaCa2+ is among the most important intracellular second
messengers participating in a plethora of biological processes, and
the measurement of Ca2+ fluctuations is significant in
the phenomenology of the underlying processes. Aequorin-based Ca2+ probes represent an invaluable tool for reliable measurement
of Ca2+ concentrations and dynamics in different subcellular
compartments. However, their use is limited due to the lack on the
market of ready-to-use, cost-effective, and portable devices for the
detection and readout of the low-intensity bioluminescence signal
produced by these probes. Silicon photomultipliers (SiPMs) are rapidly
evolving solid-state sensors for low light detection, with single
photon sensitivity and photon number resolving capability, featuring
low cost, low voltage, and compact format. Thus, they may represent
the sensors of choice for the development of such devices and, more
in general, of a new generation of multipurpose bioluminescence detectors
suitable for cell biology studies. Ideally, a detector customized
for these purposes must combine high dynamic range with high fidelity
in reconstructing the light intensity signal temporal profile. In
this article, the ability to perform aequorin-based intracellular
Ca2+ measurements using a multipurpose, low-cost setup
exploiting SiPMs as the sensors is demonstrated. SiPMs turn out to
assure performances comparable to those exhibited by a custom-designed
photomultiplier tube-based aequorinometer. Moreover, the flexibility
of SiPM-based devices might pave the way toward routinely and wide
scale application of innovative biophysical protocols.
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probephoton numberSilicon-Photomultiplier-Based Platformlow-intensity bioluminescence signalmeasurementsensorsubcellular compartmentsphoton sensitivitycell biology studiesscale applicationbioluminescence detectorssilicon photomultipliersIntracellular Calcium Dynamicsphotomultiplier tube-based aequorin...Targeted AequorinSiPM-based deviceslight detectionlight intensity signalaequorin-based intracellular
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