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Anal Chem ; 91(13): 8383-8389, 2019 Jul 02.
Artigo em Inglês | MEDLINE | ID: mdl-31140782


Intracellular pH is an important modulator of cell functions, and its subtle change may dramatically affect the cellular activities and cause diseases. A reliable imaging of the intracellular pH is still a great challenge. We imaged the intracellular pH during the cell cycle at the single living cell level using newly designed cell-penetrating peptide conjugated pH nanosensors on a home-built in situ microscopic cell culture platform. The conjugated cell-penetrating peptide greatly enhanced the uptake of nanosensors without sacrificing the pH response. We observed a gradual alkalization from interphase to prophase and rapid acidification from prometaphase to telophase, reflecting variation and consumption of the species related to the energy storage during cell cycle. We realized SERS-based pH and fluorescence dual-mode imaging when the pH sensor was further modified with fluorescence dye. The integration of SERS imaging with in situ microscopic cell culture system offers great opportunity for revealing the intracellular pH-related biological and pathological processes.

Chem Rev ; 118(10): 4946-4980, 2018 05 23.
Artigo em Inglês | MEDLINE | ID: mdl-29638112


Surface-enhanced Raman spectroscopy (SERS) inherits the rich chemical fingerprint information on Raman spectroscopy and gains sensitivity by plasmon-enhanced excitation and scattering. In particular, most Raman peaks have a narrow width suitable for multiplex analysis, and the measurements can be conveniently made under ambient and aqueous conditions. These merits make SERS a very promising technique for studying complex biological systems, and SERS has attracted increasing interest in biorelated analysis. However, there are still great challenges that need to be addressed until it can be widely accepted by the biorelated communities, answer interesting biological questions, and solve fatal clinical problems. SERS applications in bioanalysis involve the complex interactions of plasmonic nanomaterials with biological systems and their environments. The reliability becomes the key issue of bioanalytical SERS in order to extract meaningful information from SERS data. This review provides a comprehensive overview of bioanalytical SERS with the main focus on the reliability issue. We first introduce the mechanism of SERS to guide the design of reliable SERS experiments with high detection sensitivity. We then introduce the current understanding of the interaction of nanomaterials with biological systems, mainly living cells, to guide the design of functionalized SERS nanoparticles for target detection. We further introduce the current status of label-free (direct) and labeled (indirect) SERS detections, for systems from biomolecules, to pathogens, to living cells, and we discuss the potential interferences from experimental design, measurement conditions, and data analysis. In the end, we give an outlook of the key challenges in bioanalytical SERS, including reproducibility, sensitivity, and spatial and time resolution.

Materiais Biocompatíveis/análise , DNA/análise , Nanoestruturas/análise , Proteínas/análise , Análise Espectral Raman/normas , Técnicas Biossensoriais , Humanos , Propriedades de Superfície
Artigo em Inglês | MEDLINE | ID: mdl-29395932


To achieve an insightful look within biomolecular processes on the cellular level, the development of diseases as well as the reliable detection of metabolites and pathogens, a modern analytical tool is needed that is highly sensitive, molecular-specific and exhibits fast detection. Surface-enhanced Raman spectroscopy (SERS) is known to meet these requirements and, within this review article, the recent progress of label-free SERS in biological and biomedical applications is summarized and discussed. This includes the detection of biomolecules such as metabolites, nucleic acids and proteins. Further, the characterization and identification of microorganisms has been achieved by label-free SERS-based approaches. Eukaryotic cells can be characterized by SERS in order to gain information about the outer cell wall or to detect intracellular molecules and metabolites. The potential of SERS for medically relevant detection schemes is emphasized by the label-free detection of tissue, the investigation of body fluids as well as applications for therapeutic and illicit drug monitoring. The review article is concluded with an evaluation of the recent progress and current challenges in order to highlight the direction of label-free SERS in the future.

Técnicas Biossensoriais/métodos , Nanopartículas Metálicas/química , Ácidos Nucleicos/análise , Proteínas/análise , Análise Espectral Raman/métodos , Humanos , Propriedades de Superfície
J Am Chem Soc ; 137(8): 2828-31, 2015 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-25671760


Constructing nanoparticles into well-defined structures at mesoscale and larger to create novel functional materials remains a challenge. Inspired by atomic epitaxial growth, we propose an "epitaxial assembly" method to form two-dimensional nanoparticle arrays (2D NAs) directly onto desired materials. As an illustration, we employ a series of surfactant-capped nanoparticles as the "artificial atoms" and layered hybrid perovskite (LHP) materials as the substrates and obtain 2D NAs in a large area with few defects. This method is universal for nanoparticles with different shapes, sizes, and compositions and for LHP substrates with different metallic cores. Raman spectroscopic and X-ray diffraction data support our hypothesis of epitaxial assembly. The novel method offers new insights into the controllable assembly of complex functional materials and may push the development of materials science at the mesoscale.

Anal Chem ; 86(24): 12250-7, 2014 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-25418952


Local microenvironment pH sensing is one of the key parameters for the understanding of many biological processes. As a noninvasive and high sensitive technique, surface-enhanced Raman spectroscopy (SERS) has attracted considerable interest in the detection of the local pH of live cells. We herein develop a facile way to prepare Au-(4-MPy)-BSA (AMB) pH nanosensor. The 4-MPy (4-mercaptopyridine) was used as the pH sensing molecule. The modification of the nanoparticles with BSA not only provides a high sensitive response to pH changes ranging from pH 4.0 to 9.0 but also exhibits a high sensitivity and good biocompatibility, stability, and reliability in various solutions (including the solutions of high ionic strength or with complex composition such as the cell culture medium), both in the aggregation state or after long-term storage. The AMB pH nanosensor shows great advantages for reliable intracellular pH analysis and has been successfully used to monitor the pH distribution of live cells and can address the grand challenges in SERS-based pH sensing for practical biological applications.

Concentração de Íons de Hidrogênio , Nanopartículas , Soroalbumina Bovina/química , Análise Espectral Raman
Anal Chem ; 86(4): 2238-45, 2014 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-24460183


Proteins perform vital functional and structural duties in living systems, and the in-depth investigation of protein in its native state is one of the most important challenges in the postgenomic era. Surface-enhanced Raman spectroscopy (SERS) can provide the intrinsic fingerprint information of samples with ultrahigh sensitivity but suffers from the reproducibility and reliability issues. In this paper, we proposed an iodide-modified Ag nanoparticles method (Ag IMNPs) for label-free detection of proteins. The silver nanoparticles provide the huge enhancement to boost the Raman signal of proteins, and the coated iodide layer offers a barrier to prevent the direct interaction between the proteins and the metal surface, helping to keep the native structures of proteins. With this method, highly reproducible and high-quality SERS signals of five typical proteins (lysozyme, avidin, bovine serum albumin, cytochrome c, and hemoglobin) have been obtained, and the SERS features of the proteins without chromophore were almost identical to the respective normal Raman spectra. This unique feature allows the qualitative identification of them by simply taking the intensity ratio of the Raman peaks of tryptophan to phenylalanine residues. We further demonstrated that the method can also be used for label-free multiplex analysis of protein mixture as well as to study the dynamic process of protein damage stimulated by hydrogen peroxide. This method proves to be very promising for further applications in proteomics and biomedical research.

Iodetos/química , Nanopartículas/química , Proteínas/análise , Análise Espectral Raman/métodos , Animais , Bovinos , Galinhas , Hemoglobinas/análise , Muramidase/análise , Soroalbumina Bovina/análise