ArticleMolecules (Basel, Switzerland)2014
Photobleaching kinetics and time-integrated emission of fluorescent probes in cellular membranes.
Article in Molecules (Basel, Switzerland), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 54 citations in OpenAlex.
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- Plasmonic Enhancement of Fluorescence and Protein Dynamics in Living Mammalian Cells.Advanced materials (Deerfield Beach, Fla.) · 2026Article
- Dark-State-Mediated Photobleaching in mCherry-Based Red Fluorescent Proteins.The journal of physical chemistry letters · 2026Article
- Live-cell single-molecule dynamics of eukaryotic RNA polymerase machineries.Science (New York, N.Y.) · 2026Article
- Photobleaching-based autofluorescence mapping reveals distinct patterns in sporadic basal cell carcinoma, nevoid basal cell carcinoma syndrome-associated basal cell carcinoma and squamous cell carcinoma.Frontiers in medicine · 2026Article
- PerfectlyAverage: A classical open-source software method to determine the optimal averaging parameters in laser scanning fluorescence microscopy.Journal of microscopy · 2025Article
- Impact of triplet state population on GFP-type fluorescence and photobleaching.Biology of the cell · 2025Article
- Photobleaching alters the morphometric analysis of fluorescently labeled neurons and microglial cells.Pathology oncology research : POR · 2025Article
- Dual-Wavelength On-Chip Integrated Metalens for Epi-Fluorescence Single-Molecule Sensing.Sensors (Basel, Switzerland) · 2024Article
- Hour-Long, Kilohertz Sampling Rate Three-Dimensional Single-Virus Tracking in Live Cells Enabled by StayGold Fluorescent Protein Fusions.The journal of physical chemistry. B · 2024Article
- Hour-long, Kilohertz Sampling Rate 3D Single-virus Tracking in Live Cells Enabled by StayGold Fluorescent Protein Fusions.bioRxiv : the preprint server for biology · 2024Article
- Kinetics of Structural Transitions Induced by Sodium Dodecyl Sulfate in α-Chymotrypsin.ACS omega · 2023Article
- Detecting and differentiating neurotransmitters using ultraviolet plasmonic engineered native fluorescence.RSC advances · 2023Article
- Using mechanistic models and machine learning to design single-color multiplexed nascent chain tracking experiments.Frontiers in cell and developmental biology · 2023Article
- Image segmentation and separation of spectrally similar dyes in fluorescence microscopy by dynamic mode decomposition of photobleaching kinetics.BMC bioinformatics · 2022Article
- Designing a Small Fluorescent Inhibitor to Investigate Soluble Epoxide Hydrolase Engagement in Living Cells.ACS medicinal chemistry letters · 2022Article
- Dynamic Mode Decomposition of Fluorescence Loss in Photobleaching Microscopy Data for Model-Free Analysis of Protein Transport and Aggregation in Living Cells.Sensors (Basel, Switzerland) · 2022Article
- Article
- Probing chemotaxis activity in Escherichia coli using fluorescent protein fusions.Scientific reports · 2019Article
- Differential equation methods for simulation of GFP kinetics in non-steady state experiments.Molecular biology of the cell · 2018Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Since the pioneering work of Hirschfeld, it is known that time-integrated emission (TiEm) of a fluorophore is independent of fluorescence quantum yield and illumination intensity. Practical implementation of this important result for determining exact probe distribution in living cells is often hampered by the presence of autofluorescence. Using kinetic modelling of photobleaching combined with pixel-wise bleach rate fitting of decay models with an updated plugin to the ImageJ program, it is shown that the TiEm of a fluorophore in living cells can be determined exactly from the product of bleaching amplitude and time constant. This applies to mono-exponential bleaching from the first excited singlet and/or triplet state and to multi-exponential combinations of such processes. The TiEm can be used to correct for illumination shading and background autofluorescence without the need for fluorescent test layers or separate imaging of non-stained cells. We apply the method to simulated images and to images of cells, whose membranes were labelled with fluorescent sterols and sphingolipids. Our bleaching model can be extended to include a probability density function (PDF) of intrinsic bleach rate constants with a memory kernel. This approach results in a time-dependent bleach rate coefficient and is exemplified for fluorescent sterols in restricted intracellular environments, like lipid droplets. We show that for small deviations from the classical exponential bleaching, the TiEm of decay functions with rate coefficients remains largely independent of fluorescence lifetime and illumination, and thereby represents a faithful measure of probe distribution.
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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.