A systematic study on the discrepancy of fluorescence properties between in solutions and in cells: super-bright, environment-insensitive benzocoumarin dyes

Reo, Ye Jin; Jun, Yong Woong; Cho, Seo Won; Jeon, Jinseong; Roh, Hajung; Singha, Subhankar; Dai, Mingchong; Sarkar, Sourav; Kim, Hye Rim; Kim, Sojeong; Jin, Yusung; Jung, Yun Lim; Yang, Yun Jae; Joo, Jinmyoung; Ahn, Kyo Han

doi:10.1039/d0cc03586f

Cited by 30 publications

(17 citation statements)

References 18 publications

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“…Note that the quantum yield of QRP3 in pH 7.4 PBS [containing 0.1 M KCl and 0.007% BSA] is less than 1%; however, this does not necessarily mean that it is poorly emissive in the cellular environment. As we revealed recently, , the emission behavior of organic dyes, in particular dipolar dyes, in solution can be totally different from that in the cellular environment. Indeed, QRP3 provided bright fluorescence images inside the cell (see below).…”

Section: Resultsmentioning

confidence: 99%

NAD(P)H Quinone Oxidoreductase-1 in Organ and Tumor Tissues: Distinct Activity Levels Observed with a Benzo-rosol-Based Dual-Excitation and Dual-Emission Probe

et al. 2021

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NAD(P)H quinone oxidoreductase-1 (NQO1), a protective enzyme against cellular oxidative stress, is expressed abnormally high in solid tumors and thus recognized as a cancer biomarker. To develop a fluorescent NQO1 probe with practicality, we investigated benzo-rosol fluorophores linked with a known self-immolative quinone substrate. Four probe candidates exhibited ratiometric sensing behavior toward the enzyme, satisfying our orbital mismatch stratagem proposed before, under dual-excitation and dual-emission conditions that alleviate the spectral overlap issue commonly observed with the ratiometric probes based on intramolecular charge-transfer change. Among the candidates, two ester-linked compounds exhibited hydrolytic instability to water or an esterase, discouraging us to develop such ester-linked probes. One ether-linked, hydrolytically stable probe provided brighter cellular fluorescence than the other and thus was applied to ratiometric imaging of NQO1 in cells and tissues. We found that the enzyme activity levels are much different in organ tissues: stomach (56), kidney ( 22), colon (9.8), testis (7.8), bladder (5.6), lung (1.2), and muscle (1.0). Furthermore, a markedly high enzyme level (14.6-fold) was observed in a xenograft tumor tissue compared with that in a normal tissue, which suggests that such an NQO1 probe is promising for cancer diagnosis and for studying the enzyme-associated biology.

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Section: Resultsmentioning

confidence: 99%

NAD(P)H Quinone Oxidoreductase-1 in Organ and Tumor Tissues: Distinct Activity Levels Observed with a Benzo-rosol-Based Dual-Excitation and Dual-Emission Probe

et al. 2021

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“…Although the probe has a low quantum yield, it provides bright cellular fluorescence images. Such cell‐solution discrepancy is not uncommon for dipolar fluorophores, considering that the cellular environment is the third space for organic fluorophores [12] …”

Section: Resultsmentioning

confidence: 99%

A Two‐Photon Ratiometric Sensing Platform Based on Solid‐State Luminescent Benzocoumarin: Application to Prolonged Bioimaging of Hydrogen Peroxide

Kim

Sarkar

Ahn

2022

Chemistry An Asian Journal

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Fluorescent probes provide essential tools for studying biological systems. For the prolonged imaging of cellular analytes, the fast clearance of small‐molecular probes and products is a matter of concern in the quantitative analysis. Activatable probes that produce insoluble products inside cells can be used for prolonged imaging, but those with ratiometric imaging capability are rare. We disclose a novel sensing platform that is capable of prolonged imaging, in addition to ratiometric signaling for reliable quantitative analysis. Specifically, 3‐(pyridin‐4‐yl)‐8‐(pyrrolidin‐1‐yl)‐2H‐benzo[g]chromen‐2‐one and its pyridinium salt as a product/probe couple constitute the ratiometric sensing platform. As the former dye produces highly emissive insoluble nanoaggregates inside cells, a fluorescent probe in the latter form enables prolonged imaging of the target analyte in cells as well as in tissue by two‐photon microscopy.

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“…[41] In addition, we observed very high photostability in the dye (Figure S14), which is auseful property for imaging carried out under high light intensity.Given that impaired mtDNArepair activity in the brain has been implicated in aging and neurodegenerative disorders, [42] visualizing mtDNAr epair activity in the brain has the potential to be ap owerful tool in the study of these pathologies.N os uch imaging is currently available.M easuring an analyte in tissue level using af luorescent probe, however, can be easily biased by numerous factors affecting fluorescence intensity of the probe by the heterogeneous nature of tissue. [43] In order to compensate for factors that cause local variations in fluorescence signal such as microenvironment and local concentration, we adopted an ew procedure,i nw hich acute brain slices are hemi-sectioned, then one hemisphere is incubated with UBER under oxidative stress,w hile the other hemisphere is incubated with UBER under normal tissue-supportive conditions (Figure 5a). Direct comparison of these two mirror-image hemispheres compensates for non-specific fluorescence that disturbs the fluorescence signal from mtDNArepair activity,and allowed us to reliably measure the dynamic fluorescence enhancement due to glycosylation activity.…”

Section: Angewandte Chemiementioning

confidence: 99%

Fluorescence Imaging of Mitochondrial DNA Base Excision Repair Reveals Dynamics of Oxidative Stress Responses

et al. 2021

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Mitochondrial function in cells declines with aging and with neurodegeneration, due in large part to accumulated mutations in mitochondrial DNA (mtDNA) that arise from deficient DNA repair. However, measuring this repair activity is challenging. We employ a molecular approach for visualizing mitochondrial base excision repair (BER) activity in situ by use of a fluorescent probe (UBER) that reacts rapidly with AP sites resulting from BER activity. Administering the probe to cultured cells revealed signals that were localized to mitochondria, enabling selective observation of mtDNA BER intermediates. The probe showed elevated DNA repair activity under oxidative stress, and responded to suppression of glycosylase activity. Furthermore, the probe illuminated the time lag between the initiation of oxidative stress and the initial step of BER. Absence of MTH1 in cells resulted in elevated demand for BER activity upon extended oxidative stress, while the absence of OGG1 activity limited glycosylation capacity.

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A systematic study on the discrepancy of fluorescence properties between in solutions and in cells: super-bright, environment-insensitive benzocoumarin dyes

Abstract: The benzocoumarin dyes fluoresce negligibly in aqueous media but very strongly in cell, whereas representative conventional dyes display contrasting behaviour; the distinct emission behaviour of the fluorophores between in solution,...

Cited by 30 publications

References 18 publications

NAD(P)H Quinone Oxidoreductase-1 in Organ and Tumor Tissues: Distinct Activity Levels Observed with a Benzo-rosol-Based Dual-Excitation and Dual-Emission Probe

NAD(P)H Quinone Oxidoreductase-1 in Organ and Tumor Tissues: Distinct Activity Levels Observed with a Benzo-rosol-Based Dual-Excitation and Dual-Emission Probe

A Two‐Photon Ratiometric Sensing Platform Based on Solid‐State Luminescent Benzocoumarin: Application to Prolonged Bioimaging of Hydrogen Peroxide

Fluorescence Imaging of Mitochondrial DNA Base Excision Repair Reveals Dynamics of Oxidative Stress Responses

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