Nanoprobes, Inc. | Date: 2010-11-24
Described herein are a class of metal oligomers and polymers that contain both metals and organic groups. Said oligomers and polymers have utility in many applications including biomedical imaging, radiation therapy, drug delivery, and in vitro analytical techniques, such as fluorescence and phosphorescence.
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: STTR | Phase: Phase I | Award Amount: 225.00K | Year: 2016
DESCRIPTION provided by applicant We propose new probes for correlative super resolution fluorescence and electron microscopy that use biorthogonal reactions Click SNAP and HALO tags to label targets in living cells To minimize quenching of the fluorophores by the gold particles the small Undecagold gold atoms gold cluster label will be used this has minimal absorption at wavelengths above nm and therefore minimal overlap with emission of longer wavelength fluorophores Undecagold will be conjugated to ATTO N and SiR siliconized rhodamine longer wavelength fluorophores which are optimized for super resolution imaging and to Click and HALO tag ligands for bioorthogonal reactivity in living cells as well as for general labeling use We hypothesize that this probe configuration will provide much brighter fluorescence signals and will enable live cell and super resolution imaging which often photon are limiting These probes will combine high resolution dense and potentially quantitative labeling with a much smaller probe size than antibodies thus facilitating cellular delivery diffusion and targeting in living cells Preparations will be optimized to provide retenton of native fluorescence by quantum yield with average of or more fluorophores per gold label and Undecagold targeting group retention of native Click and HALO reactivity will be demonstrated by conjugation to test proteins bearing the conjugate tags followed chromatographic separation spectroscopic characterization and in vitro blot and light microscopy labeling The new probes will then be validated in correlative super resolution fluorescence and EM labeling experiments SIM STED and EM studies to a localize G protein coupled receptors GPCRs e g smoothened and b visualize the structure of the ciliary pocket by correlative fluorescence and electron microscopy Fluorescence brightness and quantum yield will be compared with those of combined fluorescent and gold immunoprobes fluorescence SIM and STED signals will be correlated with EM localization of targets PUBLIC HEALTH RELEVANCE A new class of combined fluorescent and gold labeling reagents will be developed that will enable labeling for both super resolution fluorescence microscopy and electron microscopy in a single labeling procedure furthermore these new probes will use biorthogonal reactions for labeling and thus may be used in living cells These will be the first combined fluorescent and gold labeling reagents Synthetic strategies and choice of labeling components will be optimized to minimize fluorescence quenching and deliver brighter fluorescent signals using smaller probes capable of high density labeling and fast tissue penetration Deliverables will include research tools to leverage the power of correlated electron microscopy to provide a morphological context and macromolecular localization for super resolution fluorescence microscopy to bring a new level of resolution to the study of biological processes at high resolution in both prepared specimens and in living cells In addition to the specific application proposed for validation our approach is intended to provide an enabling technology that will stimulate the development of complementary tools for other large scale projects such as the BRAIN initiative to map the structure function connectivity and plasticity of neural circuits Such systems pose great challenges for conventional microscopic methods because information is required on both the distribution of targets within entire systems and the precise localization of specific functional components at nanometer scale resolution
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 245.98K | Year: 2016
DESCRIPTION provided by applicant Every year more than people in the U S are diagnosed with primary brain tumors with deaths The five year survival rate is Primary brain tumors also account for of the cancers in children More effective treatments of malignant brain tumors are desperately needed We have pioneered the use of gold nanoparticles AuNPs to enhance radiotherapy In previous work we showed that Nanogold enhanced radiosurgery of a very aggressive orthotopic glioma in mice resulted in long term survival andgt days compared to with radiosurgery alone The AuNPs were IV administered Concerns about translation to humans of this very promising result include a the high cost of gold actually less than many antibody therapies b slow body clearance and c effectiveness of the Enhanced Permeability andamp Retention EPR effect for delivery in humans Upon further analysis we now propose a novel procedure to overcome not just these objections but the true barriers to effective glioma therapy After urgent surgery to remove the primary brain tumor and relieve dangerous intracranial pressure recurrence invariably develops Surgery typically does not remove the entire tumor especially difficult since tumor cells migrate even up to cm A number of studies have shown escaping tumor cells migrate in the peritumor edema often along white matter tracts Our hypothesis is that AuNPs can be designed to also move and distribute in this peritumor edema When infused in the primary tumor site they will because of their smaller size andquot catch upandquot to and engulf migrating tumor cells A gold concentration will be administered that will lead to enhancing radiotherapy by a factor of boosting Gy to Gy Normal brain would be spared since the X ray generated electrons from the gold only travel several microns thus tightly confining the boosted radiation dose To test this F tumor cells will be transduced with red fluorescent protein and luciferase and tumors orthotopically grown in rats The small AuNPs will be visualized with silver enhancement and the coincidence of AuNPs and tumor cells will be studied histologically Edema will be stained with anti albumin FITC Acceptable toxicity will be sought Radiotherapy will test efficacy Another advantage of this approach is that radiation resistant cancer stem cell as well as dormant and drug resistant tumor cells will be killed since the target of the increased radiation is the tumor cellsandapos DNA itself rather than any particular metabolic pathway This approach overcomes concerns of cost of an IV injection of gold a much smaller amount needed locally Loading other organs and whole body retention will be minimal and the gold no longer has to pass through a leaky endothelium it is administered directly to the edema in which the tumor cells reside Importantly it addresses the main reason why all current GBM therapies fail ineffective treatment of residual and migrant cells This approach represents a major paradigm shift for radiotherapy since a larger brain volume to cover the escaped cells will be irradiated a procedure now forbidden due to whole brain radiation constraints but now made possible via the very specific gold boost PUBLIC HEALTH RELEVANCE A new method is proposed using gold nanoparticles to kill brain tumor cells that currently evade surgery radiotherapy and chemotherapy This could significantly improve outcomes from one of the most dangerous and difficult to treat cancers
Nanoprobes, Inc. | Date: 2011-03-25
The present disclosure relates to the product, process, and use of 5 nm Nickel-Nitrilotriacetic acid (Ni-NTA) gold nanoparticles. Applications include diagnostic tests, imaging, therapies, detection technologies, gold conjugation to other molecules, and novel material constructs.
Agency: Department of Health and Human Services | Branch: National Institutes of Health | Program: SBIR | Phase: Phase I | Award Amount: 224.54K | Year: 2015
DESCRIPTION provided by applicant We have discovered a novel method to eliminate specimen charging during scanning electron microscopic examination of tissue specimens using a metallographic staining technique that imparts bulk conductivity to embedded tissue This allows increased beam exposure and dwell time and when used in conjunction with backscatter electron detection and gold nanoparticle labeling potentially affords increased resolution from current limits of around nm to as little as nm The reagent comprises a novel combination of heavy metal staining and targeted enzyme mediated metal deposition enzyme metallography or EnzMet These reagents and procedure will be investigated step by step in order to establish a mechanism for the formation of conductivity establish which reagents are required and simplify and optimize the reagents and procedure for use with other stains and labels Systematic omission of processing steps will be used to identify the critical reactions systematic omission of reagents will then be used to determine which reagents are essential Controlled variation in reaction conditions time temperature buffers and concentrations will then be conducted for each reagent in order to infer its mechanism and mode of action Once optimized the new staining methodology will be combined with gold labeling using progressively smaller gold nanoparticle probes from to nm in size These studies will be used to determine a minimum gold nanoparticle size that may be visualized within large volume samples using FIB SEM and b extent of penetration of probes into samples up to m or more in all dimensions and cutoff sizes for gold nanoparticle conjugates that allow complete penetration In addition multiple labeling will be pursued using different sized gold nanoparticle labels to differentiate pre and post synaptic proteins Connexin and Connexin respectively in the spinal cord of the Western Mosquitofish Gambusia affinis while simultaneously contouring and segmenting neuronal boundaries using the optimized conductive metallographic staining PUBLIC HEALTH RELEVANCE A new specimen preparation method and reagents will be developed that may improve the resolution of the electron microscopic analysis of large volume specimens such as entire neuronal circuits by serial section methods such as Serial Block Face scanning electron microscopy SBFSEM and Focussed Ion Beam scanning electron microscopy FIB SEM from its current limits of around nm to as little as nm This will be co developed with small gold probes that will enable the macromolecular localization of functional components such as proteins with structures such as gap junctions that are too small to be resolved by current methods This will provide a breakthrough in understanding the structure function and distribution of components of systems such as neuronal networks Deliverables will include research tools to bring a new level of resolution to large scale projects such as the BRAIN initiative to map the structure function connectivity and plasticity of neural circuits but will also be applicable to many larger organs and systems in general This poses great challenges to conventional microscopic methods because integrative neuroscience requires information on both the distribution of targets within entire neural circuits and the precise localization of specific functional components of synapses and gap junctions at nanometer scale resolution Our approach is intended to provide an enabling technology that will stimulate the development of other complementary tools for brain mapping and large scale microscopic investigation such as novel correlative instrumentation and probes data acquisition and analysis and imaging technologies