R&D at Nanoprobes, Inc.:
Milestones in Product Development

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Undecagold Labeling Identifies Firbrinogen Cross-Linking Site

The following two papers were presented at the Microscopy & Microanalysis 2010 meeting:

  1. Large Covalently Linked Fluorescent and Gold Nanoparticle Immunoprobes
    Using FluoroNanogold™ with larger gold: This study presents some new results.
  2. Gold Labeling of Protein Fusion Tags for EM
    New results using 5 nm Ni-NTA-Nanogold, and other investigations into new expression tags.

In Vivo Vascular Casting
Highly tolerated gold particles are potential X-ray contrast agents for vascular imaging and angiography. This paper, presented at Microscopy & Microanalysis 2005, describes the use of gold nanoparticles for a novel method of mappling vasculature and blood supply that may be used in vivo.

Enzymatic Metallography: A Simple and Powerful New Staining Method
Enzyme metallography is a simple new method for immunohistochemical and in situ hybridization staining and detection, in which a targeted enzyme conjugate is used to deposit metal from solution, giving a dense, black, highly resolved signal for brightfield light microscopy and for electron microscopy (papers presented at Microscopy and Microanalysis Meetings in 2002, 2004, and 2006).

5 nm Gold-Ni-NTA binds His Tags
Nanogold probes incorporating the nitrilotriacetic acid (NTA) nickel (II) chelate bind strongly and selectively to polyhistidine-tagged proteins. We now present a preliminary report of a larger 5 nm gold probe targeted in the same manner by an attached NTA-Ni(II) group (paper presented at Microscopy & Microanalysis 2005).

Nickel-NTA-Nanogold Binds his-Tagged Proteins
Gold probes targeted with the nitrilotriacetic acid (NTA) nickel (II) chelate bind strongly and selectively to polyhistidine-tagged proteins. This novel probe is much smaller than a conventional immunogold antibody conjugate and potentially useful for a wide range of applications (paper presented at Microscopy & Microanalysis 2002).

DNA Nanowires
DNA was used as the template for the deposition of positively charged Nanogold®; the gold particles were then enlarged and connected by the deposition of further gold selectively from solution using gold enhancement. The principle may be applied to 2- and 3-dimensional wiring, using hybridization of the terminal sequences to make connections. (presented at Microscopy & Microanalysis 2001).

Combined Fluorescent and Gold Probes
Six extended abstracts, presented at Microscopy Society of America (MSA) or Microscopy & Microanalysis Meetings between 1994 and 2003, describing the preparation, development and testing of new FluoroNanogold and other combined fluorescent and gold probes, are reproduced here.

Gold-Based Autometallography
A new autometallographic process has been developed by Nanoprobes which deposits gold rather than silver. See the results in our paper presented at the 1999 Annual Meeting of the Microscopy Society of America; or check the features of our new Goldenhance reagent which was used to obtain the results.

In Situ Hybridization with Nanogold®-Streptavidin
Highly sensitive, highly specific staining of target nucleotide sequences in tissue sections. When combined with catalytic reporter deposition (CARD, also known as TSA), single-copy sensitivity may be obtained for both DNA and RNA targets (protocols).

Combined Cy3 / Nanogold® Probes for Immunocytochemistry and In Situ Hybridization
Using CARD, single-copy sensitivity was obtained by both fluorescence and light microscopy.

Nanogold® Staining on Gels
Nanogold®-labeled proteins may be separated by gel electrophoresis, then visualized with high sensitivity right in the gel using LI Silver.

Preparation of Gold-Labeled Liposomes
This extended abstract from the 1996 Annual Meeting of the Microscopy Society of America gives details for using Nanogold®-labeled lipids to prepare gold-labeled liposomes and shows the structures which may be obtained.

Nanogold® Labeling of Peptides
Nanogold® may be conjugated readily to peptides at specific residues. This newsletter describes the results of three groups who used Nanogold® labeling reagents to label peptides.

RNA Labeling with Nanogold®
Monoamino-Nanogold® may be conjugated to RNA via the oxidized 2', 3' hydroxyls of the sugar. A complete reaction protocol is given.

This extended abstract, presented at the annual meeting of the Electron Microscopy Society of America in 1991, is the first published description of Nanogold® conjugates, their uses and advantages.

NanoVan Negative Stain
NanoVan is a lighter negative stain, formulated using vanadium instead of lead or uranium, which gives highly uniform staining. As a result, it is more suitable for use with Nanogold® reagents than conventional negative stains.

Conducting Polymer Films as EM Substrates
Polymer thin films prepared using substituted pyrroles (available from Nanoprobes) are conductive, extremely uniform and hydrophilic, properties which make them useful as EM substrates. This extended abstract from the XIIth International Congress on Electron Microscopy describes their preparation and use.



Under development:

Aldehyde Gold Clusters for Molecular Labeling.
Abstract from the 1995 Annual Meeting of the Microscopy Society of America, describing the preparation and use of polyaldehyde-functionalized probes. These are stable for extended periods in aqueous solution, and permit straightforward labeling of primary amines in biological molecules.

Gold Cluster Crystals
Highly purified preparations of large gold clusters can form ordered, protocrystalline arrays. Scanning Transmission Electron Microscopy results show this phenomenon in preparations of "Greengold," a large gold cluster compound similar to Nanogold® (paper presented at Microscopy & Microanalysis 2000).

Larger Covalent Gold.
Extending the advantages of Nanogold® to larger gold probes is an important goal of our research. These papers, presented at Microscopy & Microanalysis Meetings in 1999 and 2005, describe the preparation and use of novel, covalently linked larger gold conjugates.

Combined Enzymatic and Gold Probes
Combined enzymatic and gold probes, used with fluorescent or fluorogenic enzyme substrates, provide the means for correlative fluorescent and larger gold labeling, and when used with conventional organic chromogens, for combined brightfield light and electron microscopic labeling. This paper, presented at Microscopy and Microanalysis 2007, describes the preparation and use of combined horseradish peroxidase and Nanogold, 5 or 10 nm gold probes.

High Z Metal Carbonyls for Imaging and Microspectroscopy
Tetrariridium carbonyl clusters, partialy exchanged with modified tris (aryl) phosphine ligands, are not only very small, high-resolution probes for electron microscopy, but also imaging reagents for infrared and Raman microspectroscopy. Here, we describe their potential application as correlative EM and infrared microspectroscopy probes (paper presented at Microscopy & Microanalysis 2005).



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