A new molecular conjugation method has been developed to label biomolecules with optically stable metalorganic luminophores, such as tris(2,2'-bipyridyl)dichlororuthenium(II) hexahydrate (Rubpy), which are otherwise not possible for direct linking with the biomolecules. Unique biochemical properties of the biomolecule can, thus, be associated with photostable luminophores. This opens a general way to conjugate desired biomolecules using a sensitive signal transduction method. It also promotes the application of excellent luminescent materials, especially those based on photostable metalorganic luminophores, in biochemical analysis and biomolecular interaction studies. The conjugation method is based on uniform luminophore-doped silica (LDS) nanoparticles (63 +/- 4 nm). These nanoparticles have been prepared using a water-in-oil (W/O) microemulsion method. The controlled hydrolysis of tetraethyl orthosilicate (TEOS) in W/O microemulsion leads to the formation of monodisperse LDS nanoparticles. The luminophores are doped inside the nanoparticles, and the particle's silica surfaces can be used to covalently bind with biomolecules. The luminophores are well-protected from the environmental oxygen when they are doped inside the silica network. As an example, we used an antibody for leukemia cell recognition. The antibody was first immobilized onto the luminophore-doped nanoparticle through silica chemistry and then was used for leukemia cell identification by an optical microscopy imaging technique. The leukemia cells were identified easily, clearly, and with high efficiency using these antibody-coated nanoparticles. The advantages of using small, uniform luminophore-doped nanoparticles are discussed.

A water-in-oil microemulsion method has been applied for the preparation of silica-coated iron oxide nanoparticles. Three different nonionic surfactants (Triton X-100, Igepal CO-520, and Brij-97) have been used for the preparation of microemulsions, and their effects on the particle size, crystallinity, and the magnetic properties have been studied. The iron oxide nanoparticles are formed by the coprecipitation reaction of ferrous and ferric salts with inorganic bases. A strong base, NaOH, and a comparatively mild base, NH4OH, have been used in each surfactant to observe whether the basicity has some influence on the crystallization process during particle formation. Transmission electron microscopy, X-ray electron diffraction, and superconducting quantum interference device magnetometry have been employed to study both uncoated and silica-coated iron oxide nanoparticles. All these particles show magnetic behavior close to that of superparamagnetic materials. By use of this method, magnetic nanoparticles as small as 1-2 nm and of very uniform size (percentage standard deviation is less than 10%) have been synthesized. A uniform silica coating as thin as 1 nm encapsulating the bare nanoparticles is formed by the base-catalyzed hydrolysis and the polymerization reaction of tetraethyl orthosilicate in microemulsion. All experimental results are also compared with those for particles synthesized in pure water.

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In this report, we demonstrate the biochemical modification of silica based nanoparticles. Both pure and dye-doped silica nanoparticles were prepared, and their surfaces were modified with enzymes and biocompatible chemical reagents that allow them to function as biosensors and biomarkers. The nanoparticles produced in this work are uniform in size with a 1.6% relative standard deviation. They have a pure silica surface and can thus be modified easily with many biomolecules for added biochemical functionality. Specifically, we have modified the nanoparticle surfaces with enzyme molecules (glutamate dehydrogenase (GDH) and lactate dehydrogenase (LDH)) and a biocompatible reagent for cell membrane staining. Experimental results show that the silica nanoparticles are a good biocompatible solid support for enzyme immobilization. The immobilized enzyme molecules on the nanoparticle surface have shown excellent enzymatic activity in their respective enzymatic reactions. The nanoparticle surface biochemical functionalization demonstrates the feasibility of using nanoparticles for biosensing and biomarking applications.