RESUMEN
A new synthesis protocol is described to obtain a CdTe decorated magnetite bifunctional nanosystem via dodecylamine (DDA) as cross linker. High resolution transmission electron microscopy (HRTEM), energy-dispersive x-ray spectroscopy (EDAX), vibrating sample magnetometry (VSM), Fourier transform infrared spectroscopy (FTIR), diffuse reflectance spectroscopy (DRS) and fluorescence microscopy are used to characterize the constitution, size, composition and physical properties of these superparamagnetic-fluorescent nanoparticles. These CdTe decorated magnetite nanoparticles were then functionalized with anti-epidermal growth factor receptor (EGFR) antibody to specifically target cells expressing this receptor. The EGFR is a transmembrane glycoprotein and is expressed on tumor cells from different tissue origins including human leukemic cell line Molt-4 cells. The magnetite-CdTe composite nanosystem is shown to perform excellently for specific selection, magnetic separation and fluorescent detection of EGFR positive Molt-4 cells from a mixed population. Flow cytometry and confocal laser scanning microscopy results show that this composite nanosystem has great potential in antibody functionalized magnetic separation and imaging of cells using cell surface receptor antibody.
Asunto(s)
Compuestos de Cadmio/química , Medios de Contraste/química , Separación Inmunomagnética/métodos , Nanopartículas de Magnetita/química , Neoplasias/diagnóstico , Telurio/química , Línea Celular Tumoral , Supervivencia Celular , Receptores ErbB/metabolismo , Humanos , Luminiscencia , Nanopartículas de Magnetita/ultraestructura , Microscopía Confocal/métodos , Espectroscopía Infrarroja por Transformada de Fourier , Termogravimetría , Difracción de Rayos XRESUMEN
Fluorescent cadmium telluride quantum dots (CdTe QDs) are an optically attractive option for bioimaging, but are known to display high cytotoxicity. Nanoparticles synthesized from chitosan, a natural biopolymer of ß 1-4 linked glucosamine, display good biocompatibility and cellular uptake. A facile, green synthetic strategy has been developed to embed green fluorescent cadmium telluride quantum dots (CdTe QDs) in biocompatible CNPs to obtain a safer preparation than 'as is' QDs. High-resolution transmission electron microscopy showed the crystal lattice corresponding to CdTe QDs embedded in CNPs while thermogravimetry confirmed their polymeric composition. Electrostatic interactions between thiol-capped QDs (4 nm, -57 mV) and CNPs (~300 nm, +38 mV) generated CdTe QDs-embedded CNPs that were stable up to three months. Further, viability of NIH3T3 mouse fibroblast cells in vitro increased in presence of QDs-embedded CNPs as compared to bare QDs. At the highest concentration (10 µg/ml), the former shows 34 and 39% increase in viability at 24 and 48 h, respectively, as compared to the latter. This shows that chitosan nanoparticles do not release the QDs up to 48 h and do not cause extended toxicity. Furthermore, hydrolytic enzymes such as lysozyme and chitinase did not degrade chitosan nanoparticles. Moreover, QDs-embedded CNPs show enhanced internalization in NIH3T3 cells as compared to bare QDs. This method offers ease of synthesis and handling of stable, luminescent, biocompatible CdTe QDs-embedded CNPs with a favorable toxicity profile and better cellular uptake with potential for bioimaging and targeted detection of cellular components.
Asunto(s)
Materiales Biocompatibles/química , Compuestos de Cadmio/química , Quitosano/química , Colorantes Fluorescentes/química , Imagen Molecular/métodos , Puntos Cuánticos/química , Telurio/química , Animales , Materiales Biocompatibles/metabolismo , Transporte Biológico , Quitosano/metabolismo , Estabilidad de Medicamentos , Colorantes Fluorescentes/metabolismo , Ratones , Células 3T3 NIH , Fenómenos Ópticos , Solubilidad , Solventes/química , Agua/químicaRESUMEN
Microchannel is basic functional component of microfluidic chip and every step-forward of its construction technique has been receiving concern all over the world. The present work describes a novel, rapid and simple fabrication technique for building 3D microchannels in poly(dimethyl siloxane) (PDMS) elastomer. These microchannels were used for rapid detection of antigens (E. coli) by quantum dot (QD) based approach. Luminescent QD (CdTe) were synthesized by aqueous method and characterized using high resolution transmission electron microscopy (HRTEM), fluorescence spectroscopy and X-ray diffraction (XRD). The QDs were functionalized with anti-E. coli antibodies for immuno-detection. The reported process allowed easier and faster method of fabrication of circular 3D micochannels and demonstrated their potential use in an immuno-biosensor device.