ArticleBiophysical journal1999
Molecular dynamics in living cells observed by fluorescence correlation spectroscopy with one- and two-photon excitation.
Article in Biophysical journal, 1999. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 186 papers.
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Who cites it
186 citing papers in PubMed, 725 citations in OpenAlex.
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- Dynamic interaction of spliceosomal snRNPs with coilin explains Cajal body characteristics.The Journal of cell biology · 2025Article
- Photobleaching Analysis of Fluorescent Proteins in Two-Photon Microscopy at High Repetition Rates.Optics communications · 2025Article
- Single molecule dynamics in a virtual cell combining a 3-dimensional matrix model with random walks.Scientific reports · 2024Article
- Review
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- More than double the fun with two-photon excitation microscopy.Communications biology · 2024Review
- Statistical analysis of the autocorrelation function in fluorescence correlation spectroscopy.Biophysical journal · 2024Article
- Deubiquitination Detection of p53 Protein in Living Cells by Fluorescence Cross-Correlation Spectroscopy.ACS omega · 2023Article
- Two-Photon Excitation Spectra of Various Fluorescent Proteins within a Broad Excitation Range.International journal of molecular sciences · 2022Article
- Visualization of in vivo protein-protein interactions in plants.Journal of experimental botany · 2022Review
- Unraveling the hidden temporal range of fast βCommunications biology · 2022Article
- Building and Using a Two-Photon Fluorescence Cross-Correlation Spectroscopy Setup Including Fluorescence Lifetime Analysis.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Nuclear envelope mechanobiology: linking the nuclear structure and function.Nucleus (Austin, Tex.) · 2021Review
- Combined FCS and PCH Analysis to Quantify Protein Dimerization in Living Cells.International journal of molecular sciences · 2021Article
- Live-cell imaging of circadian clock protein dynamics in CRISPR-generated knock-in cells.Nature communications · 2021Article
126 more citing papers are in PubMed but not listed here.
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
Multiphoton excitation (MPE) of fluorescent probes has become an attractive alternative in biological applications of laser scanning microscopy because many problems encountered in spectroscopic measurements of living tissue such as light scattering, autofluorescence, and photodamage can be reduced. The present study investigates the characteristics of two-photon excitation (2PE) in comparison with confocal one-photon excitation (1PE) for intracellular applications of fluorescence correlation spectroscopy (FCS). FCS is an attractive method of measuring molecular concentrations, mobility parameters, chemical kinetics, and fluorescence photophysics. Several FCS applications in mammalian and plant cells are outlined, to illustrate the capabilities of both 1PE and 2PE. Photophysical properties of fluorophores required for quantitative FCS in tissues are analyzed. Measurements in live cells and on cell membranes are feasible with reasonable signal-to-noise ratios, even with fluorophore concentrations as low as the single-molecule level in the sampling volume. Molecular mobilities can be measured over a wide range of characteristic time constants from approximately 10(-3) to 10(3) ms. While both excitation alternatives work well for intracellular FCS in thin preparations, 2PE can substantially improve signal quality in turbid preparations like plant cells and deep cell layers in tissue. At comparable signal levels, 2PE minimizes photobleaching in spatially restrictive cellular compartments, thereby preserving long-term signal acquisition.
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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.