ArticleAnalytical and bioanalytical chemistry2013
Elimination of autofluorescence background from fluorescence tissue images by use of time-gated detection and the AzaDiOxaTriAngulenium (ADOTA) fluorophore.
Article in Analytical and bioanalytical chemistry, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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36 citing papers in PubMed.
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- A robust method for autofluorescence-free immunofluorescence using high-speed fluorescence lifetime imaging microscopy.Scientific reports · 2025Article
- Control of the fluorescence lifetime in dye based nanoparticles.Chemical science · 2024Article
- Multi intercalators FRET enhanced detection of minute amounts of DNA.European biophysics journal : EBJ · 2023Article
- Label-free fluorescence microscopy: revisiting the opportunities with autofluorescent molecules and harmonic generations as biosensors and biomarkers for quantitative biology.Biophysical reviews · 2023Review
- Imaging and detection of long-lived fluorescence probes in presence of highly emissive and scattering background.Experimental biology and medicine (Maywood, N.J.) · 2022Review
- Identification of cell types in multiplexed in situ images by combining protein expression and spatial information using CELESTA.Nature methods · 2022Article
- Unique Properties of Surface-Functionalized Nanoparticles for Bio-Application: Functionalization Mechanisms and Importance in Application.Nanomaterials (Basel, Switzerland) · 2022Review
- DeepSpot: A deep neural network for RNA spot enhancement in single-molecule fluorescence in-situ hybridization microscopy images.Biological imaging · 2022Article
- Magnetoelectric effect: principles and applications in biology and medicine- a review.Materials today. Bio · 2021Review
- Luminescence lifetime imaging of three-dimensional biological objects.Journal of cell science · 2021Article
- A fluorescent calix[4]arene with naphthalene units at the upper rim exhibits long fluorescence emission lifetime without fluorescence quenching.RSC advances · 2021Article
- Three-Dimensional Imaging in Stem Cell-Based Researches.Frontiers in veterinary science · 2021Review
- Out-of-Phase Imaging after Optical Modulation (OPIOM) for Multiplexed Fluorescence Imaging Under Adverse Optical Conditions.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Optical Sensing and Imaging of pH Values: Spectroscopies, Materials, and Applications.Chemical reviews · 2020Article
- Lanthanide-Based Optical Probes of Biological Systems.Cell chemical biology · 2020Review
- Dynamic contrast with reversibly photoswitchable fluorescent labels for imaging living cells.Chemical science · 2020Review
- Facile autofluorescence suppression enabling tracking of single viruses in live cells.The Journal of biological chemistry · 2019Article
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10 authors.
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Abstract
Sample autofluorescence (fluorescence of inherent components of tissue and fixative-induced fluorescence) is a significant problem in direct imaging of molecular processes in biological samples. A large variety of naturally occurring fluorescent components in tissue results in broad emission that overlaps the emission of typical fluorescent dyes used for tissue labeling. In addition, autofluorescence is characterized by complex fluorescence intensity decay composed of multiple components whose lifetimes range from sub-nanoseconds to a few nanoseconds. For these reasons, the real fluorescence signal of the probe is difficult to separate from the unwanted autofluorescence. Here we present a method for reducing the autofluorescence problem by utilizing an azadioxatriangulenium (ADOTA) dye with a fluorescence lifetime of approximately 15 ns, much longer than those of most of the components of autofluorescence. A probe with such a long lifetime enables us to use time-gated intensity imaging to separate the signal of the targeting dye from the autofluorescence. We have shown experimentally that by discarding photons detected within the first 20 ns of the excitation pulse, the signal-to-background ratio is improved fivefold. This time-gating eliminates over 96 % of autofluorescence. Analysis using a variable time-gate may enable quantitative determination of the bound probe without the contributions from the background.
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