ArticleMethods and applications in fluorescence2017
Total internal reflection fluorescence anisotropy imaging microscopy: setup, calibration, and data processing for protein polymerization measurements in living cells.
Article in Methods and applications in fluorescence, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 22 citations in OpenAlex.
- Fused deep learning enables 6D single-molecule localization in polarization-resolved microscopy.Methods and applications in fluorescence · 2026Article
- Time-Resolved Fluorescence Anisotropy from Single Molecules for Characterizing Local Flexibility in Biomolecules.Journal of visualized experiments : JoVE · 2025Article
- Modulating the Kinetics of a Fluorescence Anisotropy Immunoassay Using Tracer Point Mutations to Measure Human C-Peptide Secretion On-Chip.ACS omega · 2025Article
- Multistate kinetics of the syringe-like injection mechanism of Tc toxins.Science advances · 2025Article
- Insulin C-peptide secretion on-a-chip to measure the dynamics of secretion and metabolism from individual islets.Cell reports methods · 2023Article
- Fluorescence depolarization dynamics of ionic strength sensors using time-resolved anisotropy.Biophysical journal · 2021Article
- A concept for single-shot volumetric fluorescence imaging via orthogonally polarized excitation lattices.Scientific reports · 2019Article
- 2D polarization imaging as a low-cost fluorescence method to detect α-synuclein aggregation ex vivo in models of Parkinson's disease.Communications biology · 2018Article
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Authors and funding
4 authors at 1 institution in 2 countries.
Funding
Abstract
Fluorescence anisotropy imaging microscopy (FAIM) measures the depolarization properties of fluorophores to deduce molecular changes in their environment. For successful FAIM, several design principles have to be considered and a thorough system-specific calibration protocol is paramount. One important calibration parameter is the G factor, which describes the system-induced errors for different polarization states of light. The determination and calibration of the G factor is discussed in detail in this article. We present a novel measurement strategy, which is particularly suitable for FAIM with high numerical aperture objectives operating in TIRF illumination mode. The method makes use of evanescent fields that excite the sample with a polarization direction perpendicular to the image plane. Furthermore, we have developed an ImageJ/Fiji plugin, AniCalc, for FAIM data processing. We demonstrate the capabilities of our TIRF-FAIM system by measuring [Formula: see text]-actin polymerization in human embryonic kidney cells and in retinal neurons.
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Registered trials
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