RESUMO
A novel and versatile optical reader for microfluidic platforms is presented. The reader includes a modular insertion port based on the lock and key concept for reproducible alignment with a miniaturized optical detection system comprising an interchangeable light emitting diode (LED) and a photodiode. The modular nature of the insertion port allows the use of microfluidic platforms in variable shapes and fluidic configurations. Three different analytical methodologies based on absorbance or fluorescence measurements were used to demonstrate the flexibility and reproducibility of the proposed experimental setup.
Assuntos
Colorimetria/instrumentação , Fluorometria/instrumentação , Técnicas Analíticas Microfluídicas/instrumentação , Desenho de EquipamentoRESUMO
Recognition of chemical modifications in canonical nucleobases of nucleic acids is of key importance since such modified variants act as different genetic encoders, introducing variability in the biological information contained in DNA. Herein, we demonstrate the feasibility of direct SERS in combination with chemometrics and microfluidics for the identification and relative quantification of 4 different cytosine modifications in both single- and double-stranded DNA. The minute amount of DNA required per measurement, in the sub-nanogram regime, removes the necessity of pre-amplification or enrichment steps (which are also potential sources of artificial DNA damages). These findings show great potentials for the development of fast, low-cost and high-throughput screening analytical devices capable of detecting known and unknown modifications in nucleic acids (DNA and RNA) opening new windows of activity in several fields such as biology, medicine and forensic sciences.
Assuntos
Citosina/análise , Citosina/química , DNA/química , Análise Espectral Raman , Propriedades de SuperfícieRESUMO
Efficient treatments in bacterial infections require the fast and accurate recognition of pathogens, with concentrations as low as one per milliliter in the case of septicemia. Detecting and quantifying bacteria in such low concentrations is challenging and typically demands cultures of large samples of blood (~1 milliliter) extending over 24-72 hours. This delay seriously compromises the health of patients. Here we demonstrate a fast microorganism optical detection system for the exhaustive identification and quantification of pathogens in volumes of biofluids with clinical relevance (~1 milliliter) in minutes. We drive each type of bacteria to accumulate antibody functionalized SERS-labelled silver nanoparticles. Particle aggregation on the bacteria membranes renders dense arrays of inter-particle gaps in which the Raman signal is exponentially amplified by several orders of magnitude relative to the dispersed particles. This enables a multiplex identification of the microorganisms through the molecule-specific spectral fingerprints.
Assuntos
Bactérias/química , Líquidos Corporais/microbiologia , Análise Espectral Raman , Anticorpos Antibacterianos/imunologia , Bactérias/isolamento & purificação , Escherichia coli/química , Escherichia coli/imunologia , Escherichia coli/isolamento & purificação , Humanos , Nanopartículas Metálicas/química , Microfluídica/métodos , Microscopia Eletrônica de Transmissão , Pseudomonas aeruginosa/química , Pseudomonas aeruginosa/imunologia , Pseudomonas aeruginosa/isolamento & purificação , Prata/química , Streptococcus/química , Streptococcus/imunologia , Streptococcus/isolamento & purificaçãoRESUMO
The recent needs in the nanosciences field have promoted the interest towards the development of miniaturized and highly integrated devices able to improve and automate the current processes associated with efficient nanomaterials production. Herein, a green tape based microfluidic system to perform high temperature controlled synthetic reactions of nanocrystals is presented. The device, which integrates both the microfluidics and a thermally controlled platform, was applied to the automated and continuous synthesis of CdSe quantum dots. Since temperature can be accurately regulated as required, size-controlled and reproducible quantum dots could be obtained by regulating this parameter and the molar ratio of precursors. The obtained nanocrystals were characterized by UV-vis and fluorescence spectrophotometry. The band width of the emission peaks obtained indicates a narrow size distribution of the nanocrystals, which confirms the uniform temperature profile applied for each synthetic process, being the optimum temperature at 270 °C (full width at half maximum = 40 nm). This approach allows a temperature controlled, easy, low cost and automated method to produce quantum dots in organic media, enhancing its application from laboratory-scale to pilot-line scale processes.
RESUMO
In this paper, a computer controlled microreactor to synthesize water soluble CdS and CdS/ZnS nanocrystals with in situ monitoring of the reaction progress is developed. It is based on ceramic tapes and the Low-Temperature Co-fired Ceramics technology (LTCC). As well the microsystem set-up, the microreactor fluidic design has also been thoroughly optimized. The final device is based on a hydrodynamic focusing of the reagents followed by a three-dimensional micromixer. This generates monodispersed and stable CdS and core-shell CdS/ZnS nanocrystals of 4.5 and 4.2 nm, respectively, with reproducible optical properties in terms of fluorescence emission wavelengths, bandwidth, and quantum yields, which is a key requirement for their future analytical applications. The synthetic process is also controlled in real time with the integration of an optical detection system for absorbance and fluorescence measurements based on commercial miniaturized optical components. This makes possible the efficient managing of the hydrodynamic variables to obtain the desired colloidal suspension. As a result, a simple, economic, robust and portable microsystem for the well controlled synthesis of CdS and CdS/ZnS nanocrystals is presented. Moreover, the reaction takes place in aqueous medium, thus allowing the direct modular integration of this microreactor in specific analytical microsystems, which require the use of such quantum dots as labels.