Probes For Biological Applications Research Articles

Mitochondrial targeting fluorescent probe for specific detection of nickel ions and biological imaging

Nickel ion, a kind of essential trace element, is indispensable for maintaining life health. Nickel ions in excess will induce a series of reactions in the body and be correlative with the diseases and even cancer. As one of the most important organelles, mitochondrion is responsible for maintaining the homeostasis of metal ions in cells. Therefore, sensitive detection of nickel ions in mitochondria contributes to further study the physiological function of mitochondria, and is of great significance for investigating body health. In this study, a simple and effective fluorescence probe method for the detection of nickel ions in organisms and environment was proposed. The probe has shown excellent spectral performance in vitro experiments, and the detection limits was as low as 17 nM. And it can be applied to the detection of nickel ions in environmental water samples with satisfactory results, the recovery rate is higher than 85%. More importantly, the probe can specifically target to mitochondria to realize the imaging of nickel ions in cells and provide an effective auxiliary tool for in-depth study of the relationship between mitochondrial function and metal ions. In addition, the probe has been successfully applied to the detection of nickel ions in zebrafish model, which further demonstrates the great potential and value of the probe in biological applications.

Increasing the activity of DNAzyme based on the telomeric sequence: 2’-OMe-RNA and LNA modifications

Abstract2’-OMe-RNA analogues and LNA point modifications of DNA oligonucleotides were applied for the modulation of the G-quadruplex topology and enhancement of peroxidase activity of the resulting DNAzymes. The effect of the 2’-OMe-RNA analogue was studied for full length modified oligonucleotides with various sequences. In the case of LNA-point modification, we have chosen a telomeric DNA sequence and investigated various numbers of modifications. Our main goal was to prove that the application of these modifications can influence the activity of DNAzyme, especially those, which normally form poor DNAzymes. As an example, we have chosen the telomeric HT22 sequence which is known to form DNAzyme characterized by low activity. In all cases, the DNAzymes formed by a telomeric sequence with the application of the 2’-OMe-RNA analogue as well as LNA-point modification, showed significantly higher peroxidase activity. We were also able to shift the formation of hybrid or antiparallel topology to parallel topology. These results are important for the development of probes for biological applications as well as for the design of probes based on DNA sequences that normally form DNAzymes with low activity. This paper also provides information on how the application of nucleotide analogues can transform the topology of G-quadruplexes.

On the Aggregation Behaviour and Spectroscopic Properties of Alkylated and Annelated Boron‐Dipyrromethene (BODIPY) Dyes in Aqueous Solution

AbstractThe tendency of boron‐dipyrromethene (BODIPY) dyes to associate in water is well known, and usually a cause for inferior fluorescence properties. Synthetic efforts to chemically improve BODIPYs’ water solubility and minimize this problem have been numerous in the past. However, a deeper understanding of the phenomena responsible for fluorescence quenching is still required. Commonly, the spectroscopic behaviour in aqueous media has been attributed to aggregate or excimer formation, with such works often centring on a single BODIPY family. Herein, we provide an integrating discussion including very diverse types of BODIPY dyes. Our studies revealed that even subtle structural changes can distinctly affect the association behaviour of the fluorophores in water, involving different photophysical processes. The palette of behaviour found ranges from unperturbed emission, to the formation of H or J aggregates and excimers, to the involvement of tightly bound, pre‐formed excimers. These results are a first step to a more generalized understanding of spectroscopic propertiesvs. structure, facilitating future molecular design of BODIPYs, especially as probes for biological applications.

MMP2-sensing up-conversion nanoparticle for fluorescence biosensing in head and neck cancer cells

Upconversion nanoparticles (UCNPs) have extensive biological-applications because of their bio-compatibility, tunable optical properties and their ability to be excited by infrared radiation. Matrix metalloproteinases (MMPs) play important roles in extracellular matrix remodelling; they are usually found to significantly increase during cancer progression, and these increases may lead to poor patient survival. In this study, we produced a biosensor that can be recognized by MMP2 and then be unravelled by the attached quencher to emit visible light. We used 3.5-nm gold nanoparticles as a quencher that absorbed emission from UCNPs at a wavelength of 540nm. The biosensor consists of an upconversion nanoparticle, MMP2-recognized polypeptides and quenchers. Here, UCNPs consisting of NaYF4:Yb3+/Er3+ were prepared via a high temperature co-precipitation method while protecting the oleic acid ligand. To improve the biocompatibility and modify the UCNPs with a polypeptide, they were coated with a silica shell and further conjugated with MMP-recognizing polypeptides. The polypeptide has two ends of featuring carboxylic and thiol groups that react with UCNPs and AuNPs, and the resulting nanoparticles were referred to as UCNP@p-Au. According to the in vitro cell viability analysis, UCNP@p-Au exhibited little toxicity and biocompatibility in head and neck cancer cells. Cellular uptake studies showed that the MMP-based biosensor was activated by 980-nm irradiation to emit green light. This MMP-based biosensor may serve as sensitive and specific molecular fluorescent probe in biological-applications.

5-Cyanotryptophan as an infrared probe of local hydration status of proteins

The nitrile (C N) stretching vibration is sensitive to environment, making nitrile-derivatized amino acids an increasingly utilized tool to study various biological processes. Herein, we show that the bandwidth of the C N stretching vibration of 5-cyanotryptophan is particularly sensitive to water, rendering it an attractive infrared probe of local hydration status. We confirm the utility of this probe in biological applications by using it to examine how the hydration status of individual tryptophan sidechains of an antimicrobial peptide, indolicidin, changes upon peptide binding to model membranes. Furthermore, we show that p-cyanophenylalanine and 5-cyanotryptophan constitute a useful fluorescence energy transfer pair.

Reduced blinking and long-lasting fluorescence of single fluorophores coupling to silver nanoparticles.

The fluorescence signal of single organic fluorophores is characterized by random blinking and irreversible photobleaching. Photoinduced blinking of Cy5 has posed various limitations of this popular near-infrared (NIR) probe in biological applications. Here we show that fluorophore-metal nanoparticle (NP) complexes greatly suppress Cy5 blinking and noticeably reduce photobleaching events. The blinking behavior of single Cy5 molecules was investigated and compared in the absence and the presence of silver nanostructures. A power-law distribution of off time population was observed for single Cy5 molecules. Average off times were compared to evaluate the plasmonic effect of silver nanoparticles on the triplet decay rates. We furthermore demonstrate enhanced photostability in the presence of silver NPs. The results show that plasmonic-controlled fluorescence can lead to a novel physical mechanism to enhance fluorescence intensity, reduce blinking, and increase photostability.

Biocompatible Polymeric Nanoparticles and Films Containing Hydrophobic Quantum Dots

ABSTRACTMicro- or nano-particles encapsulating hydrophobic quantum dots were prepared by the emulsion technique. Films containing quantum dots were also prepared by film casting methods. Quantum dots without hydrophilic coating were directly mixed with polymer solution to prepared extremely stable films or particles that do not phase-separate with time. The surface of the particles or films could be modified to have different hydrophilicity and/or functional groups. Particles with 1.5 ± 0.16 μm and 320 ± 26 nm in diameters and films of 300 μm in thickness were prepared. NIH 3T3 and EMT-6 were culture on the films containing quantum dots for 8 – 20 h. Compared to the control, quantum dot were delivered directly and efficiently into the cells without toxicity. When implanted near tumor cells in Balb/C mice, quantum dots migrated from the films into the tumor cells for at least 3 days. Considering that the quantum dot dispersion in aqueous media is relatively unstable and difficult to handle, our stable particles or films containing hydrophobic quantum dots can become versatile probes for biological applications.

In vitro and in vivo measurement for biological applications using micromachined probe

We developed a small-sized micromachined probe for the measurement of biological properties using microelectromechanical systems (MEMS) technology. We also experimentally showed the suitability of the micromachined probe for biological applications through in vivo, as well as in vitro measurements of various types of tissue. We measured the permittivities of 0.9% saline and the muscle and fat of pork using the micromachined probe after liquid calibration. The measured permittivities of 0.9% saline and pork agreed well with both the expected values of the Cole-Cole equation along with the measured values obtained through the use of a 1-mm-diameter open-ended coaxial probe. We also performed in vivo measurements of breast cancer tissue implanted in an athymic nude mouse to show the suitability of the small-sized micromachined probe for practical biological applications. Through the obtained data, the capability of the micromachined probe of distinguishing different tissue types from one another was shown. The actual aperture size of the micromachined probe is only 240 /spl mu/m /spl times/ 70 /spl mu/m and, therefore, we can extract the biological information from very small biological tissues and drastically decrease the invasiveness of this method through the implementation of the small probe created through the use of MEMS technology.

Europium-doped calcium pyrophosphates: Allotropic forms and photoluminescent properties

In a search for new luminescent biological probes, we synthesized calcium pyrophosphates doped with europium up to an atomic Eu/(Eu+Ca) ratio of 2%. They were prepared by coprecipitating a mixture of calcium and europium salts with phosphate. After heating at 900 °C in air, two phases coexisted, identified as the β calcium pyrophosphate form and EuPO 4. Heating near 1250 °C in air, during the β → α transformation, europium ions substitute for calcium ions in the α calcium pyrophosphate structure as demonstrated by the spectroscopic study. Europium ions with both valence states (divalent and trivalent) were observed in the samples. Following the synthesis procedure, partial reduction of Eu 3+ took place even in an oxidizing atmosphere. The 0.5%-doped compound could serve as a sensitive probe in biological applications. Depending on the excitation wavelength, the luminescence occurs either in the red or in the blue regions, which discriminates it from parasitic signals arising from other dyes or organelles in live cells.

Permittivity measurements up to 30 GHz using micromachined probe

We implemented a micromachined probe for the measurement of biological properties using MEMS technology, and experimentally showed the suitability of the micromachined probe in biological applications. The micromachined probe was fabricated on a silicon substrate, and to remove wave transmission through the silicon substrate, we etched the silicon substrate from beneath a lower ground and made the etched silicon surface conducting by using thermal evaporation of Cr/Au and a coating of conductive epoxy. The micromachined probe consists of a CPW and strip line between benzo cyclo butene (BCB) layers, which is known to be a material with high resistivity, low loss tangent, and low permittivity at high frequency. We measured the permittivity of a number of well-known liquids—0.5%, 0.9% and 1.3% saline, acetone, ethanol, and muscle and fat of pork—as biological samples using the micromachined probe after liquid calibration. The measured permittivity of 0.9% saline agreed well with the expected value of the Cole–Cole equation. In this paper, we first demonstrate that the micromachined probe can provide broadband measurement of measurable solid materials, such as biological samples, and also of well-known liquids at microwave frequencies. The size of the micromachined probe is 2000 µm (width) × 580 µm (thickness) × 30 000 µm (length), and the aperture size of the micromachined probe is only 650 µm × 70 µm. Therefore, we can extract the biological information from very small biological tissues and reduce radiation effects. Thus we show the feasibility of low-cost, small and portable permittivity measurement systems using a micromachined open-ended coaxial RF MEMS probe.

Synthesis of a Far Red Absorbing Thiobarbituric Acid Oxonol Derivative as a Fluorescent Probe for Biological Applications

A water-soluble oxonol dye containing a reactive hydroxysuccinimidyl ester suitable for reaction with biomolecules has been prepared. This fluorescent compound has 631 nm, εabs 147,000 M−1 cm−1, 659 nm, and quantum yield 0.30.

Characterization of a Series of Far Red Absorbing Perylene Diones: A New Class of Fluorescent Probes for Biological Applications

The far red region of the spectrum is increasingly being utilised in many applications in the biosciences. However, apart from the cyanine group of dyes, there are relatively few far red fluorescent probes available which are of practical use. We have synthesised and characterised a new class of far red fluorescent probes based on the perylene dione chromophore. The 2,10 di-substituted perylene diones possess broad absorption spectra (>90 nm bandwidths), large Stokes shifts (>60 nm) and quantum yields of up to 0.5 with a maximum absorption at 610–640 nm in organic solvents or in solutions of non-ionic detergents. A number of derivatives have been synthesised that can be used as membrane probes, as chromogenic substrates for alkaline phosphatase, and for the labelling of macromolecules such as proteins and DNA. These novel far red fluorophores have potential applications in diagnostic and research applications.