RESUMO
As the third-generation nucleic acid amplification technology, digital polymerase chain reaction (PCR) has been widely adopted in the analysis of nucleic acids. However, further application of this powerful technology is hindered due to the limitation of surfactants. Here, for the first time, we propose the use of colloidosomes self-assembled from fluorinated silica nanoparticle for digital PCR to address this limitation. A one-step fluorinated silica nanoparticle synthesis method is proposed, which is much more convenient and reproducible compared with the synthesis of conventional fluorine-based surfactants. Fluorinated silica nanoparticles facilitate the formation of colloidosomes with excellent stability capable of enduring the rapid temperature changes associated with the PCR and avoiding material exchange (cross-talk) between droplets for high-fidelity analysis. The colloidosome digital PCR method was developed using these colloidosomes as highly parallel reactors for single-copy nucleic acid amplification and rare mutant detection. The method is robust and accurate, and it offers possibilities for a great variety of applications, such as gene expression studies, single cell analysis, and circulating tumor DNA detection.
Assuntos
Coloides/química , Neoplasias do Colo/genética , Mutação , Nanocápsulas/química , Reação em Cadeia da Polimerase/métodos , Proteínas Proto-Oncogênicas p21(ras)/genética , Tensoativos/química , Adenocarcinoma/genética , Adenocarcinoma/patologia , Neoplasias do Colo/patologia , Análise Mutacional de DNA/métodos , Humanos , Dióxido de Silício , Células Tumorais CultivadasRESUMO
In the past decade, tremendous efforts have been devoted to the search for the alternatives to Pt-based catalysts for the oxygen reduction reaction (ORR) in fuel cells and metal-air batteries. Recently, metal-nitrogen-carbon (M-N-C) systems, especially 3d transition metals (TM) and their alloys encapsulated in nitrogen-doped carbon based materials (TM@N-C), have attracted increasing attention due to their low cost and high ORR activity. Here, a simple and novel strategy is developed to synthesize sandwich-structured TM@N-C composites, in which ultrafine Fe nanoparticles are encapsulated in nitrogen-doped carbon nanotubes (N-CNTs) grafted on both sides of reduced graphene oxide (rGO) sheets by pyrolysis of ammonium ferric citrate-functionalized zeolitic imidazolate framework-8@graphene oxide (Fe@ZIF-8@GO). The resulting Fe@N-CNTs@rGO composites naturally integrate zero-dimensional (0D) Fe nanoparticles, one-dimensional (1D) N-CNTs, and two-dimensional (2D) graphene into a three-dimensional (3D) hierarchical architecture with highly dispersed active sites, a large surface area, and abundant porosity. Because of these structural advantages, the sandwich-structured Fe@N-CNTs@rGO composites display a half-wave potential of 0.83 V in a 0.1 M KOH solution for the ORR, comparable to that of commercial Pt/C catalysts, and more excellent durability and resistance to fuel molecules. The proposed strategy paves a new way for the synthesis of non-precious high-performance electrocatalysts for energy conversion applications.