Fluorescent Probes For Bioimaging Research Articles

Large stokes shift and near-infrared fluorescent probe for bioimaging and evaluating the HClO in an rheumatoid arthritis mouse model

It is crucial to identify aberrant HClO levels in living things since they pose a major health risk and are a frequent reactive oxygen species (ROS) in living organisms. In order to detect HClO in various biological systems, we created and synthesized a near-infrared fluorescent probe with an oxime group (−C = N-OH) as a recognition unit. The probe DCMP1 has the advantages of fast response (10 min), near-infrared emission (660 nm), large Stokes shift (170 nm) and high selectivity. This probe DCMP1 not only detects endogenous HClO in living cells, but also enables further fluorescence detection of HClO in living zebrafish. More importantly, it can also be used for fluorescence imaging of HClO in an rheumatoid arthritis mouse model. This fluorescent probe DCMP1 is anticipated to be an effective tool for researching HClO.

Clarifying the role of carboxyl functionality in squaraines towards aggregation and self-assembly through photophysical characterization and BSA interactions

The demand for near-infrared (NIR) probes with robust and stable fluorescence signals has surged, driven by their applications in optical sensing and bioimaging. Squraine dyes as NIR fluorescent probes have gained significant traction. However, understanding their behavior in aqueous media is important for their successful application. Herein we report the synthesis and photophysical characterization of carboxy-functionalized unsymmetrical squaraine dyes capable of light absorption and emission in the near-infrared (NIR) wavelength region. Structural variations in alkyl chain length, and spacer chain length between the primary chromophore and –COOH group were systematically introduced to investigate their impact on self-assembly in aqueous media leading to dye aggregation and their differential interactions with Bovine Serum Albumin (BSA) as a model protein. In phosphate buffer (PB) the absorption intensity decreased drastically along with pronounced shoulders due to dye aggregation leading to complete fluorescence quenching. However, in the presence of BSA, the intensity of shoulder bands reduced coupled with a sharp increase in the absorption associated with monomeric dye with the increasing concentration of BSA, indicating the prevention of dye aggregation owing to dye-BSA interactions. The prevention of dye aggregation due to dye-BSA interaction also led to a tremendous increase in the fluorescence intensities as a function of the increasing BSA concentrations. Site-selective study off BSA showed that these dyes bind preferably at Site-II using hydrophilic interactions. Thus, these dyes show great potential as FRET or fluorescent on/off probes for biosensing, non-covalent labeling of proteins, and fluorescent probes for bioimaging.

Advances in the application of biosynthesized carbon dots as fluorescent probes for bioimaging

Advances in the application of biosynthesized carbon dots as fluorescent probes for bioimaging

Can a Small Change in the Heterocyclic Substituent Significantly Impact the Physicochemical and Biological Properties of (Z)-2-(5-Benzylidene-4-oxo-2-thioxothiazolidin-3-yl)acetic Acid Derivatives?

Rhodanine-3-acetic acid derivatives are attractive compounds with versatile effects. What is very important is that compounds of this type have many biological properties. They are tested, among others, as fluorescent probes for bioimaging and aldose reductase inhibitors. Rhodanine-3-acetic acid derivatives also have antibacterial, antifungal and anticancer activity. The presented work demonstrates that a slight change in the five-membered heterocyclic substituent significantly affects the properties of the compounds under consideration. Three rhodanine-3-acetic acid derivatives (A-1-A-3) were obtained in the Knoevenagel condensation reaction with good yields, ranging from 54% to 71%. High thermal stability of the tested compounds was also demonstrated above 240 °C. The absorption and emission maxima in polar and non-polar solvents were determined. Then, the possibility of using the considered derivatives for fluorescence bioimaging was checked. Compounds A-1 and A-2 were successfully used as fluorescent dyes of fixed cells of mammalian origin. In addition, biological activity tests against bacteria and fungi were carried out. Our results showed that A-1 and A-2 showed the most excellent antimicrobial activity among the newly synthesized compounds, especially against Gram-positive bacteria.

A near-infrared emitting “off-on” fluorescent probe for bioimaging of Pd(Ⅱ) ions in living cells and mice

BackgroundThe surging consumption of palladium in modern industry has given rise to its accumulation in the ecosystem, posing conspicuous toxicity to aquatic organisms and human health. The investigation of palladium in biological systems is highly demanded for the in-depth understanding of its dynamics and behaviors. Fluorescence imaging serves as a powerful approach to assess palladium species in biological systems, and currently most of the sensing probes are applicable to living cells. Effective tracking of palladium species in living organisms is challenging, which requires sufficient hydrophilicity and imaging depth of the probes. ResultsBased on an intramolecular charge transfer (ICT) mechanism, a distyryl boron dipyrromethene (BODIPY) derivative (DISBDP-Pd) has been prepared for the near-infrared (NIR) fluorescence imaging of Pd2+ ions. Two additional methoxy triethylene glycol (TEG) chains could serve as flexible and hydrophilic moieties to enhance the aqueous solubility and cell permeability of the extended conjugate. Solution studies revealed that DISBDP-Pd exhibited a NIR fluorescence enhancement signal exclusively to Pd2+ ions (detection limit as low as 0.85 ppb) with negligible interference from Pd0 species and other closely related metal ions. Computational calculations have been performed to rationalize the binding mode and the mechanism of action. Fluorescence imaging assays have been conducted on A549 human non-small cell lung carcinoma cells and mouse models. Exhibiting negligible cytotoxicity, DISBDP-Pd demonstrated concentration-related fluorescence enhancement signals in response to Pd2+ ions in living cells and mice. SignificanceDISBDP-Pd exhibits advantages over many small molecule palladium probes in terms of satisfactory aqueous solubility, high sensitivity and selectivity, and biocompatible NIR emission property, which are particularly favorable for the sensing application in biological environments. The design strategy of this probe can potentially be adopted for the functionalization of other BODIPY probes implemented for NIR fluorescence bioimaging.

A novel dual near-infrared fluorescent probe for bioimaging and visualization of viscosity in acute alcoholic liver injury.

A dual NIR fluorescent probe Cy-ND is developed for viscosity sensing with λex/em = 766/806 nm, making it apt for biological analysis, whose response is validated through DFT and TDDFT computations. Cy-ND successfully detected viscosity changes amidst acute alcohol-induced liver injury and liver ischemia-reperfusion injury.

Azide-modified corrole phosphorus complexes for endoplasmic reticulum-targeted fluorescence bioimaging and effective cancer photodynamic therapy

Study on corrole photosensitizers (PSs) for photodynamic therapy (PDT) has made remarkable progress. Targeted delivery of PSs is of great significance for enhancing therapeutic efficiency, decreasing the dosage, and reducing systemic toxicity during PDT. The development of PSs that can be specifically delivered to the subcellular organelle is still an attractive and challenging work. Herein, we synthesize a series of azide-modified corrole phosphorus and gallium complex PSs, in which phosphorus corrole 2-P could not only precisely target the endoplasmic reticulum (ER) with a Pearson correlation coefficient (PCC) up to 0.92 but also possesses the highest singlet oxygen quantum yields (ΦΔ = 0.75). This renders it remarkable PDT activity at a very low dosage (IC50 = 23 nM) towards HepG2 tumor cell line while ablating solid tumors in vivo with excellent biosecurity. Furthermore, 2-P exhibits intense red fluorescence (ΦF = 0.25), outstanding photostability, and a large Stokes shift (190 nm), making it a promising fluorescent probe for ER. This study provides a clinically potential photosensitizer for cancer photodynamic therapy and a promising ER fluorescent probe for bioimaging.

A fluorescent probe for bioimaging of GSH in cancer cells

A fluorescent probe CTP2-IMC was designed for bioimaging of glutathione (GSH) in cancer cells with indomethacin (IMC), coumarin and bromide as the targeting group, fluorophore and receptor, respectively. Due to the π-π interaction between coumarin and IMC, CTP2-IMC mainly exists in the form of folded state in aqueous solution. The non-radiative transitions caused by the photo-induced electron transfer (PET) process from IMC to the fluorophore as well as the heavy-atom effect led to non-fluorescent of CTP2-IMC. The substitution of Br by GSH and unfolded conformation induced by IMC acceptor on cancer cells resulted in significant fluorescence enhancement, which enabled CTP2-IMC to bioimage GSH in cancer cells rather than in normal one.

Reaction-Activated Disassembly of the NIR-II Probe Enables Fast Detection and Ratiometric Photoacoustic Imaging of Glutathione In Vivo.

Glutathione (GSH), the most abundant nonprotein biothiol, is a significant endogenous molecule that plays a key role in redox equilibrium in vivo and is regarded as a critical biomarker of cancer. Currently, various fluorescent probes have been designed and synthesized for imaging GSH at the cellular level in the visible range and the first near-infrared window (NIR-I, 750-900 nm). However, the application of these fluorescent probes for bioimaging and biosensing in vivo has been extremely hindered by the high biobackground and low tissue penetration. Herein, based on the self-assembly and disassembly of J-aggregation, we designed and synthesized a GSH-activatable probe MC-PSE for second near-infrared window (NIR-II) fluorescence and ratiometric photoacoustic imaging of GSH in vivo. The anionic cyanine-based MC-PSE tends to form stable J-aggregates in an aqueous solution. Upon the reaction with GSH, the J-aggregates of MC-PSE disassembled, the emission peak intensity of MC-PSE at 940 nm significantly increased by about 20 times, and the PA900/PA980 ratio increased by 4 times within 15 min in vitro. Notably, we used MC-PSE to visualize GSH in tumor-bearing mice and to distinguish normal and tumor areas successfully by virtue of NIR-II FL and PA dual-modal imaging. The design strategy of MC-PSE provides a novel method for ratiometric photoacoustic imaging, and MC-PSE is expected to be a powerful tool for the accurate detection of GSH in cancer diagnosis.

Amphiphilic Fluorinated Unimer Micelles as Nanocarriers of Fluorescent Probes for Bioimaging.

The unique self-assembly properties of unimer micelles are exploited for the preparation of fluorescent nanocarriers embedding hydrophobic fluorophores. Unimer micelles are constituted by a (meth)acrylate copolymer with oligoethyleneglycol and perflurohexylethyl side chains (PEGMA90-co-FA10) in which the hydrophilic and hydrophobic comonomers are statistically distributed along the polymeric backbone. Thanks to hydrophobic interactions in water, the amphiphilic copolymer forms small nanoparticles (<10 nm), with tunable properties and functionality. An easy procedure for the encapsulation of a small hydrophobic molecule (C153 fluorophore) within unimer micelles is presented. UV-vis, fluorescence, and fluorescence anisotropy spectroscopic experimental data demonstrate that the fluorophore is effectively embedded in the nanocarriers. Moreover, the nanocarrier positively contributes to preserve the good emissive properties of the fluorophore in water. The efficacy of the dye-loaded nanocarrier as a fluorescent probe is tested in two-photon imaging of thick ex vivo porcine scleral tissue.

An Efficient Green Approach to Constructing Adenine Sulfate-Derived Multicolor Sulfur- and Nitrogen-Codoped Carbon Dots and Their Bioimaging Applications.

A cost-effective and environmentally friendly approach is proposed for producing N- and S-codoped multicolor-emission carbon dots (N- and S-codoped MCDs) at a mild reaction temperature (150 °C) and relatively short time (3 h). In this process, adenine sulfate acts as a novel precursor and doping agent, effectively reacting with other reagents such as citric acid, para-aminosalicylic acid, and ortho-phenylenediamine, even during solvent-free pyrolysis. The distinctive structures of reagents lead to the increased amount of graphitic nitrogen and sulfur doping in the N- and S-codoped MCDs. Notably, the obtained N- and S-codoped MCDs exhibit considerable fluorescence intensities, and their emission color can be adjusted from blue to yellow. The observed tunable photoluminescence can be attributed to variations in the surface state and the amount of N and S contents. Furthermore, due to the favorable optical properties, good water solubility and biocompatibility, and low cytotoxicity, these N- and S-codoped MCDs, especially green carbon dots, are successfully applied as fluorescent probes for bioimaging. The affordable and environmentally friendly synthesis method employed to create N- and S-codoped MCDs, combined with their remarkable optical properties, offers a promising avenue for their use in various fields, particularly in biomedical applications.

Single and two-photo excited violet-blue fluorescence from aza [6]helicenes and the plannar precursors based-on indole, carbazole, and fluorene

Organic π-conjugated materials exhibiting violet-blue emissions are of great interest for diverse applications. Helicenes and stilbenes hold great potential as compounds capable of achieving violet-blue fluorescence. In this study, aza[6]helicenes THNI and DHNC, along with their planar precursors DFVHI and HHIVC, were successfully synthesized using indole, carbazole, and fluorene units as building blocks. The molecular structures were verified through the utilization of 1H and 13C nuclear magnetic resonance (NMR), high resolution mass spectroscopies (HRMS), and the crystal structure of THNI was further elucidated via X-ray analysis. The synthesized material exhibited exceptional thermal stability with a Td value of ≥350 °C. The HOMO and LUMO energy levels were determined through cyclic voltammetry analysis, while density functional theory (DFT) and time-dependent DFT (TD-DFT) calculations at the B3LYP/6-31G(d) level were employed to investigate ground-state geometries and vertical electronic transitions. Optical properties were characterized using UV–vis absorption spectroscopy as well as single- and two-photon excited photoluminescence. The UV–vis absorption spectra indicate that helicenes possess superior light-harvesting capabilities compared to their planar precursors, owing to the former's higher molar extinction coefficients. These compounds exhibit violet-blue emissions in various solvents and solid-state films. In addition, aza[6]helicenes exhibit higher photoluminescence quantum yields (PLQYs) and shorter maximum emission peaks compared to their planar precursors. However, the maximum two-photon absorption (TPA) cross section (δ) values of aza[6]helicenes are lower than those of their planar counterparts. Therefore, THNI and DHNC may be promising candidates for violet-blue emitters in OLEDs due to their curved confined π-electron systems, high thermal and optical stability, and high PLQYs. Meanwhile, DFVHI and HHIVC exhibit potential as two-photon fluorescent probes for bioimaging.

Endogenous formaldehyde responsive fluorescent probe for bioimaging

Endogenous formaldehyde responsive fluorescent probe for bioimaging

One-Pot Three Component Synthesis of Quinazolin-4(3H)-One Derivatives: Investigation of Photophysical Properties and FRET Application toward Protein Lysozyme

A one-pot, three-component reaction between isatoic anhydride, 1,3-diketones, and amines/arylhydrazines catalyzed by a combination of molecular I2 and Et3N in open-air affords a series of quinazolin-4(3H)-one derivatives in good yield. The protocol offers an operationally simple, facile, and cost-effective pathway utilizing easily accessible reagents under mild reaction conditions with significant substrate scope. The results of steady state and time-resolved absorption and emission spectroscopy indicate the potential of quinazolin-4(3H)-one derivatives as a fluorescence probe. They exhibit unique photophysical responses in different solvent environments. In addition, they can also act as FRET acceptors during interaction with protein lysozyme. Preliminary biological studies indicate that quinazolin-4(3H)-ones are cell permeable and can be used as a fluorescent probe for bio-imaging of human cervical cancer (HeLa) cells.

A highly selective and sensitive red-emitting fluorescent probe for bioimaging of hypobromous acid in living cells

Hypobromous acid (HOBr) is an indispensable reactive oxygen species which is necessary for the functioning of many cellular functions. However, abnormal HOBr can cause serious harm to the environment and organisms. Nowadays, fluorescent probe techniques are widely used in the detection of biomolecules in living bodies, but few fluorescent probes have been investigated for the detection of HOBr in living cells. Based on the above considerations, we synthesized a fluorescent probe RD-HOBr for detecting HOBr. Probe RD-HOBr has the characteristics of fast response (<4 s), high selectivity and naked eye detectability. Notably, because the detection limit of the probe RD-HOBr for detecting HOBr is only 1.37 nM, it has the advantage of high sensitivity to HOBr. In addition, the probe RD-HOBr can not only detect HOBr in actual water samples, but also has the advantage of low toxicity and can detect HOBr in cells sensitively. Therefore, the construction of probe RD-HOBr not only provides a new tool for the sensitive detection of HOBr, but is also expected to provide a new method for the detection of HOBr in biological organisms.

New cationic spiropyrans with photoswitchable NIR fluorescence

Visible-light-mediated photochromic compounds with NIR absorption and fluorescence are of great interest for use in different biomedical applications. In this work, new representatives of spiropyrans with conjugated cationic 3H-indolium substituents in different positions of 2H-chromene moiety were synthesized. The electron-donating methoxy groups were introduced in the uncharged indoline and charged indolium cycles to form the effective conjugation chain between the hetarene moiety and the cationic fragment for reaching NIR absorption and fluorescence. The molecular structure and the effects of cationic fragment position on the mutual stability of the spirocyclic and merocyanine forms of compounds were carefully studied in the solutions and solid state by NMR, IR, HRMS, single-crystal XRD, and quantum chemical calculations. It was found that the obtained spiropyrans demonstrate positive or negative photochromism depending on the cationic fragment’s position. One of spiropyrans has shown bidirectional photochromic properties induced exclusively by visible light of different wavelengths in both directions. The photoinduced merocyanine forms of compounds possessed far-red shifted absorption maxima and NIR fluorescence, which makes them prospective fluorescent probes for bioimaging.

New High-Performance Fluorescent Dye Scaffolds: Applications for Bioimaging and Biosensing

AbstractFluorescence imaging in the shortwave infrared region (SWIR, 850–2500 nm) window has become an indispensable tool in biomedical research because it has weaker absorption, less light scattering, and less background fluorescence in this window. As we know, the design and synthesis of fluorescent dyes and fluorescent probes are the cores of fluorescence imaging and biosensing. Thus, their photophysical mechanisms exploring and bioanalytical applications are multidisciplinary and cutting-edge research topics. In this regard, we have been working on high-performance fluorescent dyes and fluorescent probes for years. In short, we have developed a series of bright, stable, aggregation-resistant, SWIR fluorescent dyes ECXs, based on a carbon-bridged spiro ring strategy. We also developed a series of high-performance SWIR fluorescent dyes CXs and Chrodols, which combine the structural advantages of cyanine and xanthenoid dyes. Based on these new SWIR scaffolds, we further constructed some activatable SWIR fluorescent probes with OFF-ON or ratiometric properties for biosensing in vivo. Therefore, the main line of our work is to gain an in-depth understanding of the photophysical mechanisms of fluorescent dyes, to create high-performance luminescent dyes, and to further develop fluorescent probes for bioimaging and biosensing.1 Introduction2 ECX Dyes Based on a Carbon-Bridged Spiro Ring Strategy3 Fluorescent Dyes and Fluorescent probes Combining the Structures of Cyanine and Xanthenoid Dyes3.1 CX Series Dyes3.2 A Functionalized Modified CX probe NRh3.3 CX-like Dyes with a Secondary Amino Py-23.4 Chrodol Series Dyes4 Conclusion and Future Prospects

Hyaluronic acid-magnetic fluorescent polydopamine/gold-based fluorescent probe for bioimaging of CD44 over-expressed cancer cells

CD44 is a transmembrane glycoprotein (P-glycoprotein) that expressed in a large scale on the surface membrane of breast cancer cells. The CD44 receptor has been employed for various medial aims such as imaging, drug delivery, and etc. In the present work, the surface of citrate-capped gold nanoparticles (cit-AuNPs) was modified with magnetic polydopamine nanoparticles to provide fluorescent nanocomposite and amine groups for attachment to hyaluronic acid (HA) molecules. HA molecules have been bonded to the surface of fluorescent polydopamine nanoparticles (FPNPs) via the formation of amide bonds between carboxyl (COOH) group of HA and amine groups of the FPNPs. Due to the presence of the HA molecules, the MNPs-FPNPs/AuNPs/HA can attach to the CD44-overexpressed MCF-7 cells with high specificity and could be employed for bioimaging aims. Fluorescence microscopy was applied for bioimaging of the MNPs-FPNPs/AuNPs/HA internalized cancer cells. Obtained results offered that the MNPs-FPNPs/AuNPs/HA can especially and efficiently internalize to the cells and differentiate CD44-positive cancer cells and normal cells of HFFF-2 cells. Also, the MNPs-FPNPs/AuNPs/HA has a biocompatible nature with the toxic levels around 250 μg/mL levels. As a conclusion, the MNPs-FPNPs/AuNPs/HA is a good candidate for specific bioimaging applications of CD44−overexpressed MCF-7 cancer cells with high physicochemical stability and advantageous spectral features.

Biogenic preparation of undoped and heteroatoms doped carbon dots: effect of heteroatoms doping in fluorescence, catalytic ability and multicolour in-vitro bio-imaging applications - A comparative study

The proficient carbon nanocatalysts and incredible fluorescent bio-imaging probes have been successfully fabricated and compared their remarkable efficiencies in this work. The work describes the preparation of undoped and heteroatoms (N, B & S) doped carbon dots from the Syzygium malaccense (S. malaccense) fruit extract via a simple and low cost hydrothermal carbonization method. The obtained pure and heteroatoms doped CDs were characterized and confirmed spectrometrically through various analytical techniques. The size of the obtained CDs, NCDs, B-CDs and S-CDs were found to be 4.68, 4.50, 3.71 and 3.10 nm, respectively. HR-TEM studies explored that the synthesized CDs were spherical in shape and the size of the CDs was reduced during doping of heteroatoms. The photophysical properties of the dissimilar CDs were enlightened through UV–Vis spectroscopy and spectrofluorometer. The quantum yield (QY) was found to be 0.32, 0.41, 0.35 and 0.36 for synthesized CDs, NCDs, B-CDs and S-CDs, respectively. The XRD, Raman and SAED results exposed the amorphous nature of the heteroatoms doped CDs. FT-IR, EDAX and XPS studies helped to confirm the doping of heteroatoms (N, B and S) were doped over the CDs surface. The proficient catalytic ability of the various CDs was revealed and compared towards the methylene blue (MB) reduction using NaBH4. Among the synthesized CDs, the NCDs showed better catalytic ability and efficiently reduced the MB within 10 min. Furthermore, the fluorescent staining ability of the prepared CDs was examined and compared in the in-vitro multicolour HeLa cell imaging. The NCDs were found to give high intense HeLa cell imaging compared to other synthesized CDs.

Recent Advance in Nucleus-Targeted Fluorescent Probes for Bioimaging, Detection and Therapy

The nucleus is not only the control hub of cell life activities, but also the center of storing and controlling genetic information. The morphology of the nucleus can be used to judge the cell vitality and health. The various biological molecules contained in the nucleus are closely related to the normal life activities, occurrence and development of diseases. In recent years, fluorescence imaging has gained increasing attention due to its advantages of being intuitive, in situ and visual. The development of fluorescent probes for high-resolution imaging of the nucleus and detection of biomolecules in the nucleus is of great value for understanding the normal physiological processes of cells or organisms, as well as for disease diagnosis and treatment. In this review, we mainly introduce the current general strategies for designing nucleus-targeted fluorescent probes, as well as their applications in nucleus-targeted imaging, intranuclear biomolecular detection and therapy. Moreover, we also discuss the current challenges and opportunities of nucleus-targeted fluorescent probes in terms of selectivity, design strategies and so on.