ArticleBio-protocol2019
Triple Fluorescence Anisotropy Reporter Imaging in Living Cells.
Article in Bio-protocol, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.
Who cites it
4 citing papers in PubMed, 8 citations in OpenAlex.
- A Novel Single-Color FRET Sensor for Rho-Kinase Reveals Calcium-Dependent Activation of RhoA and ROCK.Sensors (Basel, Switzerland) · 2024Article
- Comparative CaBiophysical reports · 2021Article
- PIE-FLIM Measurements of Two Different FRET-Based Biosensor Activities in the Same Living Cells.Biophysical journal · 2020Article
- Article
Corrections and comments
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
Abstract
FRET-based genetically encoded biosensors incorporate two fluorescent proteins into their design to enable ratiometric biosensing of signaling activities in live cells. While emission ratios are generally useful for quantitative studies, they leave little room in the optical spectrum for additional sensors and optogenetic tools. Homotransfer-based reporters, such as the FLuorescence Anisotropy REporters (FLAREs), incorporate two fluorescent proteins of the same color into their design. Conversion to a single color opens the visible spectrum for the use of complementary sensors. Here, we present a protocol for measuring three independent intracellular signals in living cells. We describe the configuration and calibration of a widefield microscope for multicolor FLARE imaging. Three FLARE sensors for intracellular calcium, MAPK activity, and PKA phosphorylation are co-transfected into HEK293 cells, and triple FRET imaging is performed. Compared to heterotransfer FRET biosensors, the polarization-based multiplex imaging can track multiple signaling activities concurrently in a targeted cell population.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.