Ratiometric pH sensing by fluorescence resonance energy transfer-based hybrid semiconducting polymer dots in living cells

Fluorescent microscopy has enabled the study of intracellular processes and revealed the most intricate details of the subcellular structure. This has benefitted not only the basic biological science, but also has had an impact in numerous biomedical applications. Basic fluorescent sensing techniques use the change in the absolute emission of a fluorescent sensor. This entails some disadvantages as the signal might be influenced by factors not directly related to the process under study (e.g., fluctuations in the excitation source). To overcome these drawbacks, one can use multiple emissions of a single or various fluorophores. There are numerous examples of multichannel fluorescence microscopy techniques that have given rise to numerous ratiometric methods and multiplexing assays. Herein, how the use of multiple emission channels has impacted fluorescence microscopy in terms of speed, sensitivity, and resolution is reviewed. Using recent examples, how the easy implementation of multichannel detection can overcome current limitations of the main used fluorescence techniques and promote the development of novel microscopy methods is shown.

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“…These techniques can make use of the strengths of multichannel methods if the emission of the donor is also exploited. [ 79,94,95 ]…”

Section: Sensing Techniquesmentioning

confidence: 99%

Multichannel Fluorescence Microscopy: Advantages of Going beyond a Single Emission

Rodríguez‐Sevilla

Thompson

Jaque

2022

Advanced NanoBiomed Research

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“…[1] Fluorescent materials, ranging from molecular dyes to emissive nanoparticles, exhibit appealing optical and chemical properties, which allow them to be ideal fluorophores. [2][3][4][5] Upon most occasions, sole fluorophore is inadequate to achieve the goal of "lighting up" a specific biological component/event unless a carrier is incorporated in the system. The carriers can provide a sequence of different advanced effects, relying Cellulose nanocrystals (CNCs) are featured with low toxicity, non-trivial biocompatibility, and cell membrane penetration capability, which allow the constructions of nanoplatforms for biosensing and in vivo imaging.…”

Section: Introductionmentioning

confidence: 99%

Design and Construction of Fluorescent Cellulose Nanocrystals for Biomedical Applications

Liu

Seidi

et al. 2022

Adv Materials Inter

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Cellulose nanocrystals (CNCs) are featured with low toxicity, non‐trivial biocompatibility, and cell membrane penetration capability, which allow the constructions of nanoplatforms for biosensing and in vivo imaging. Interfacing CNCs and fluorescent materials into sensors/probes is thus highly topical and has received tremendous interest. This review covers the development of CNC extraction methods and, in particular, their impacts on the surficial properties of CNCs. Whereafter, recently reported strategies for fluorescent functionalizations of CNCs are summarized based on chemical modification, physical adsorption, or in situ growth. Choosing the most suitable strategy, according to the properties of both CNCs and fluorophores, for constructing fluorescent CNCs is also emphasized. With regard to applications of the fluorescent CNCs, this work focuses on the studies which have involved but not been limited to metal ion sensing, physiological pH detection, cell imaging, and tumor antiproliferation. Being aware of the highly flexible construction, appealing structural/optical properties, and outstanding performances in analysis/imaging, it is believed that CNCs are bound to be increasingly investigated in the future and widely applied in the biomedical area.

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“…Semiconducting polymer dots (Pdots) are fluorescent nanoparticles [14][15][16][17] that exhibit superior photophysical properties relative to small fluorophores,i norganic semiconducting quantum dots,a nd fluorescent proteins.P dots exhibit high brightness,f ast emission, large absorption cross-section, and high photostability,a sw ell as low toxicity and versatile surface modification, [18][19][20][21] making them well-suited for applications in cell labeling, [22] in vivo imaging, [23,24] single-particle tracking, [25] drug/gene delivery, [26] and tumor therapy. [27] Pdots also have been used as biosensors [28,29] to detect pH, [30,31] temperature, [32] metal ions, [33] oxygen, [34] glucose, [35,36] and exosomes. [37] Here we design as eries of NADH-responsive Pdots for use as an NADH sensor,a nd characterize the Pdots in vitro and in vivo.N ADH sensing is achieved by electron transfer from aP dot to NADH, resulting in quenching of Pdot fluorescence emission.…”

Section: Introductionmentioning

confidence: 99%

Reversible Ratiometric NADH Sensing Using Semiconducting Polymer Dots

Chen

Men

et al. 2021

Angew Chem Int Ed

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Reduced nicotinamide adenine dinucleotide (NADH) is a key coenzyme in living cells due to its role as an electron carrier in redox reactions, and its concentration is an important indicator of cell metabolic state. Abnormal NADH levels are associated with age‐related metabolic diseases and neurodegenerative disorders, creating a demand for a simple, rapid analytical method for point‐of‐care NADH sensing. Here we develop a series of NADH‐sensitive semiconducting polymer dots (Pdots) as nanoprobes for NADH measurement, and test their performance in vitro and in vivo. NADH sensing is based on electron transfer from semiconducting polymer chains in the Pdot to NADH upon UV excitation, quenching Pdot fluorescence emission. In polyfluorene‐based Pdots, this mechanism resulted in an on‐off NADH sensor; in DPA‐CNPPV Pdots, UV excitation resulted in NADH‐sensitive emission at two wavelengths, enabling ratiometric detection. Ratiometric NADH detection using DPA‐CNPPV Pdots exhibits high sensitivity (3.1 μM limit of detection), excellent selectivity versus other analytes, reversibility, and a fast response (less than 5 s). We demonstrate applications of the ratiometric NADH‐sensing Pdots including smartphone‐based NADH imaging for point‐of‐care use.

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Ratiometric pH sensing by fluorescence resonance energy transfer-based hybrid semiconducting polymer dots in living cells

Cited by 19 publications

References 34 publications

Multichannel Fluorescence Microscopy: Advantages of Going beyond a Single Emission

Multichannel Fluorescence Microscopy: Advantages of Going beyond a Single Emission

Design and Construction of Fluorescent Cellulose Nanocrystals for Biomedical Applications

Reversible Ratiometric NADH Sensing Using Semiconducting Polymer Dots

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