Effects of Viscosity and Refractive Index on the Emission and Diffusion Properties of Alexa Fluor 405 Using Fluorescence Correlation and Lifetime Spectroscopies

Zanten, Camila van; Melnikau, Dzmitry; Ryder, Alan G.

doi:10.1007/s10895-021-02719-y

Cited by 5 publications

(4 citation statements)

References 55 publications

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“…On the other hand, the solution viscosity will also greatly influence the fluorescence or scattering intensity of probes. 42 Figure S5 confirms this trend that the BPP values of AuNPs or AgNPs will decrease with the increase in viscosity. However, the experiment confirmed that the influence of the viscosity on the SBR seemed negligible, as shown in Figure 1g,h.…”

Section: ■ Results and Discussionsupporting

confidence: 65%

“…So, the uncertainty of the translational diffusion time due to the nonuniform viscosity within living cells results in that it is difficult to use the characteristic diffusion time as a quantitative parameter. On the other hand, the solution viscosity will also greatly influence the fluorescence or scattering intensity of probes Figure S5 confirms this trend that the BPP values of AuNPs or AgNPs will decrease with the increase in viscosity.…”

Section: Resultssupporting

confidence: 54%

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Brightness Analysis per Moving Particle: In Situ Analysis of Alkaline Phosphatase in Living Cells

et al. 2022

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In situ quantitative analysis of enzymes such as phosphatase is important to understand a number of involved biological processes ranging from various metabolisms to signal transduction and cellular regulation. In this paper, a novel in situ measurement strategy was proposed to detect alkaline phosphatase (ALP) activity in different locations within single living cells. The principle is based on the measurement of the resonance light scattering brightness ratio (SBR) per moving nanoparticle that forms in an ALP-related chemical reaction. In the method, a novel resonance light scattering correlation spectroscopy (RLSCS) system was developed using two lasers for illumination or two detection channels. Using the gold nanoparticles (AuNPs) as probes, the Au@Ag nanoparticles (Au@Ag NPs) formed due to the ALP-catalyzed hydrolysis of ascorbic acid 2-phosphate (AAP) and the subsequent reduction–deposition reaction of Ag ions that occurred on the AuNPs. The SBR value per moving particle was determined based on the obtained RLS intensity traces and RLSCS curves. The SBR value was found to be not influenced by the intracellular viscosity and size that was confirmed in the experiments. The linear relation between the SBR and ALP activity was established and applied to detect ALP activity and evaluate the inhibition of different drugs. Finally, the method was successfully used to in situ measure ALP activity within living cells. The method overcomes the shortcoming of conventional methods that lack quantitative analysis and are susceptible to intracellular viscosity.

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Section: ■ Results and Discussionsupporting

confidence: 65%

Section: Resultssupporting

confidence: 54%

Brightness Analysis per Moving Particle: In Situ Analysis of Alkaline Phosphatase in Living Cells

et al. 2022

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“…38 For Laurdan measurements, the lifetime calibration was obtained using Alexa 405 considering a single-exponential decay with 3.4 ns lifetime. 39 3. RESULTS…”

Section: Methodsmentioning

confidence: 99%

“…FLIM calibration of the system was performed by measuring the known lifetime of the fluorescein (for di-4-ANEPPDHQ and CF-TP10) that is a single-exponential of 4.0 ns . For Laurdan measurements, the lifetime calibration was obtained using Alexa 405 considering a single-exponential decay with 3.4 ns lifetime …”

Section: Methodsmentioning

confidence: 99%

Peptide–Membrane Interactions Monitored by Fluorescence Lifetime Imaging: A Study Case of Transportan 10

et al. 2021

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The interest on detailed analysis of peptide–membrane interactions is of great interest in both fundamental and applied sciences as these may relate to both functional and pathogenic events. Such interactions are highly dynamic and spatially heterogeneous, making the investigation of the associated phenomena highly complex. The specific properties of membranes and peptide structural details, together with environmental conditions, may determine different events at the membrane interface, which will drive the fate of the peptide–membrane system. Here, we use an experimental approach based on the combination of spectroscopy and fluorescence microscopy methods to characterize the interactions of the multifunctional amphiphilic peptide transportan 10 with model membranes. Our approach, based on the use of suitable fluorescence reporters, exploits the advantages of phasor plot analysis of fluorescence lifetime imaging microscopy measurements to highlight the molecular details of occurring membrane alterations in terms of rigidity and hydration. Simultaneously, it allows following dynamic events in real time without sample manipulation distinguishing, with high spatial resolution, whether the peptide is adsorbed to or inserted in the membrane.

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Estimating Poly(N-isopropylacrylamide) size in solution below the LCST using fluorescence correlation spectroscopy with non-covalent bound fluorophores

van Zanten,

Ryder

2024

Polymer Testing

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Effects of Viscosity and Refractive Index on the Emission and Diffusion Properties of Alexa Fluor 405 Using Fluorescence Correlation and Lifetime Spectroscopies

Cited by 5 publications

References 55 publications

Brightness Analysis per Moving Particle: In Situ Analysis of Alkaline Phosphatase in Living Cells

Brightness Analysis per Moving Particle: In Situ Analysis of Alkaline Phosphatase in Living Cells

Peptide–Membrane Interactions Monitored by Fluorescence Lifetime Imaging: A Study Case of Transportan 10

Estimating Poly(N-isopropylacrylamide) size in solution below the LCST using fluorescence correlation spectroscopy with non-covalent bound fluorophores

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