Fluorescence lifetime imaging with time-gated detection of hyaluronidase using a long lifetime azadioxatriangulenium (ADOTA) fluorophore

Chib, Rahul; Requena, Sebastian; Mummert, Mark E.; Strzhemechny, Yuri M.; Gryczyński, Ignacy; Borejdo, Julian; Gryczyński, Zygmunt; Fudala, Rafał

doi:10.1088/2050-6120/4/4/047001

Cited by 8 publications

(13 citation statements)

References 63 publications

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“…Hydrophobicity of HA can in particular emerge upon special treatments of HA (e.g., freeze-drying) or upon specific functionalization . The mix of hydrophobic patches and hydrophilic moieties allows, upon functionalization with proper dyes, to create nanostructured particles and gels in which fluorescent moieties are mainly aggregated and that, most interestingly, can be disaggregated in response to specific stimuli, in particular, under the action of HAase. ,− …”

Section: Introductionsupporting

confidence: 65%

Specific, Surface-Driven, and High-Affinity Interactions of Fluorescent Hyaluronan with PEGylated Nanomaterials

Palomba

Rampazzo

Zaccheroni

et al. 2020

ACS Appl. Mater. Interfaces

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Hybrid nanomaterials are a subject of extensive research in nanomedicine, and their clinical application is reasonably envisaged in the near future. However, the fate of nanomaterials in biological environments poses serious limitations to their application; therefore, schemes to monitor them and gain control on their toxicity could be of great help for the development of the field. Here, we propose a probe for PEGylated nanosurfaces based on hyaluronic acid (HA) functionalized with rhodamine B (RB). We show that the high-affinity interaction of this fluorogenic hyaluronan (HA-RB) with nanoparticles exposing PEGylated surfaces results in their sensing, labeling for super-resolution imaging, and synergistic cellular internalization. HA-RB forms nanogels that interact with high affinity—down to the picomolar range—with silica nanoparticles, selectively when their surface is covered by a soft and amphiphilic layer. This surface-driven interaction triggers the enhancement of the luminescence intensity of the dyes, otherwise self-quenched in HA-RB nanogels. The sensitive labeling of specific nanosurfaces also allowed us to obtain their super-resolution imaging via binding-activated localization microscopy (BALM). Finally, we show how this high-affinity interaction activates a synergistic cellular uptake of silica nanoparticles and HA-RB nanogels, followed by a differential fate of the two partner nanomaterials inside cells.

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Section: Introductionsupporting

confidence: 65%

Specific, Surface-Driven, and High-Affinity Interactions of Fluorescent Hyaluronan with PEGylated Nanomaterials

Palomba

Rampazzo

Zaccheroni

et al. 2020

ACS Appl. Mater. Interfaces

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“…63 Time-correlated single photon counting (TCSPC) technique based on SPAD or PMT detectors was used in TG imaging. 13,29,39,47 In addition, various°uorescence imaging and detection based on TG technique was also demonstrated, including°uorescence lifetime imaging (FLIM), 13,26,43,47,[64][65][66] TG F€ orster resonance energy transfer (FRET), 40,67-69 and°uorescence di®use optical tomography. 33…”

Section: Detectionmentioning

confidence: 99%

“…Further, FLIM technique is able to produce lifetime maps, which is basically implemented by gradually adjusting the TG delay. 13,26,43,47,[64][65][66] TCSPC is one of the main approaches to realize FLIM. TGFI has been widely utilized in many biological and medical applications.…”

Section: Biological Applicationsmentioning

confidence: 99%

“…For quite long luminescence lifetime (e.g., lanthanide-based probes) up to millisecond time scale, 32,71,72,74,76,78,81 simple and low-cost instrumentation (mainly optical-chopper-based methods) is well satisfactory. By contrast, for quite short luminescence lifetime down to nanosecond time scale, 28,29,31,33,34,47,66 complex and expensive instrumentation (mainly electronic-unit-based methods) must be employed.…”

Section: Biological Applicationsmentioning

confidence: 99%

“…Recovery of the concealed micro objects such as large molecules, 11,28 cells, 11,23,29,34,36,39,41,47,49,66,70,72,[74][75][76][77][78][79][80][81][82][83][84] and microorganisms 24,30,32,71,85 from annoying interference (e.g., background AF) in°uorescence imaging with the help of TG technique has attracted increasing attention for years. Marriott et al presented a simple TG microscope and utilized it to visualize the delayed luminescence intensities and lifetime of acridine dyes on polytene chromosomes.…”

Section: Molecules Cells and Microorganismsmentioning

confidence: 99%

See 2 more Smart Citations

Time-gated fluorescence imaging: Advances in technology and biological applications

Yang

Chen

2020

J. Innov. Opt. Health Sci.

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Time-gated (TG) fluorescence imaging (TGFI) has attracted increasing attention within the biological imaging community, especially during the past decade. With rapid development of light sources, image devices, and a variety of approaches for TG implementation, TGFI has demonstrated numerous biological applications ranging from molecules to tissues. The paper presents inclusive TG implementation mainly based on optical choppers and electronic units for synchronization of fluorescence excitation and emission, which also serves as guidelines for researchers to build suited TGFI systems for selected applications. Note that a special focus will be put on TG implementation based on optical choppers for TGFI of long-lived probes (lifetime range from microseconds to milliseconds). Biological applications by TG imaging of recently developed luminescent probes are described.

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Time‐Resolved Luminescent Sensing and Imaging for Enzyme Catalytic Activity Based on Responsive Probes

Wang

Jin

et al. 2022

Chemistry An Asian Journal

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Enzymes, as a kind of biomacromolecules, play an important role in many physiological processes and relate directly to various diseases. Developing an efficient detection method for enzyme activity is important to achieve early diagnosis of enzyme-relevant diseases and high throughput screening of potential enzyme-relevant drugs. Time-resolved luminescence assay provide a high accuracy and signal-tonoise ratios detection methods for enzyme activity, which has been widely used in high throughput screening of enzyme-relevant drugs and diagnosis of enzyme-relevant diseases. Inspired by these advantages, various responsive probes based on metal complexes and metal-free organic compounds have been developed for time-resolved bioimaging and biosensing of enzyme activity owing to their long luminescence lifetimes, high quantum yields and photostability. In this review, we comprehensively reviewed metal complex-and metal-free organic compound-based responsive probes applied to detect enzyme activity through timeresolved imaging, including their design strategies and sensing principles. Current challenges and future prospects in this rapidly growing field are also discussed.

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Fluorescence lifetime imaging with time-gated detection of hyaluronidase using a long lifetime azadioxatriangulenium (ADOTA) fluorophore

Cited by 8 publications

References 63 publications

Specific, Surface-Driven, and High-Affinity Interactions of Fluorescent Hyaluronan with PEGylated Nanomaterials

Specific, Surface-Driven, and High-Affinity Interactions of Fluorescent Hyaluronan with PEGylated Nanomaterials

Time-gated fluorescence imaging: Advances in technology and biological applications

Time‐Resolved Luminescent Sensing and Imaging for Enzyme Catalytic Activity Based on Responsive Probes

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