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Supersensitive in Situ Hybridization by Tyramide Signal Amplification and Nanogold Silver Staining: The Contribution of Autometallography and Catalyzed Reporter Deposition to the Rejuvenation of In Situ Hybridization (open access)

Supersensitive in Situ Hybridization by Tyramide Signal Amplification and Nanogold Silver Staining: The Contribution of Autometallography and Catalyzed Reporter Deposition to the Rejuvenation of In Situ Hybridization

It is peculiar that in situ hybridization (ISH), a technique with many similarities to immunohistochemistry (IHC), has not enjoyed the phenomenal growth in both basic research and clinical applications as has its sister technique IHC. Since the late 1970s, when immunoperoxidase techniques began to be applied to routine diagnostic material and to numerous research applications, there has been a natural evolution of the IHC procedure. Namely, only a few primary antibodies were available commercially at the onset, and only one indirect and the peroxidase-antiperoxidase (PAP) technique detection systems were in place. With the advent of avidin-biotin detection systems and monoclonal antibodies, and a viable commercial market, extraordinary growth of the procedure's applications in clinical research and diagnostic pathology occurred during the subsequent two decades. Today, IHC is automated and widely used for research purposes and, to a large extent, has become a routine diagnostic ''special stain'' in most clinical laboratories. During the same period, ISH enjoyed very little growth in both research and diagnostic applications. What has accounted for this lack of maturation of the technique? The success of IHC is part of the reason measuring a gene's encoded protein routinely and inexpensively, particularly as automation evolved, rendered IHC a …
Date: April 17, 2002
Creator: Tubbs, Raymond R.; Pettay, James; Grogan, Thomas; Powell, R. D.; Cheung, Annie L. M.; Hainfeld, James et al.
System: The UNT Digital Library

Oral History Interview with Bruce Gordon Elliot, May 17, 2002

Access: Use of this item is restricted to the UNT Community
Interview with Bruce Elliot, a Navy veteran and POW from Montezuma, Kansas. Elliot discusses his family, joining the Navy and volunteering for Asiatic service, the start of war and the bombing of Cavite Navy Yard in the Philippines, retreat to Corregidor, capture by the Japanese, escaping internment on Palawan and joining Moro guerillas, sabotage, linking up with Australian forces, evacuation to Australia and returning to the United States, becoming a deepsea diver, and Korean War service. In appendix are a photo of Elliot, a map of the Philippines, two photos of a POW camp on Palawan, and a photo of three of his comrades.
Date: May 17, 2002
Creator: Alexander, Bill & Elliot, Bruce Gordon
System: The UNT Digital Library
Silver and Gold Based Autometallography of Nanogold. (open access)

Silver and Gold Based Autometallography of Nanogold.

For many applications, silver salt-based autometallography (often also called silver enhancement or silver development) is required to visualize colloidal gold (1-5 nm in diameter) or the small 1.4 nm Nanogold{reg_sign} particles (Nanoprobes, Yaphank, NY, USA). Although even Nanogold may be seen directly by scanning-transmission electron microscopy (STEM), by transmission EM (TEM; in thin sections without stain or ice-embedded cryo-EM samples), energy filtered TEM, and scanning EM (SEM), silver enhancement makes viewing in the EM more facile since the particles are enlarged to approximately 10 to 20 nm, convenient for most specimens. Autometallographic (AMG) enhancement is required in order to visualize smaller gold particles such as Nanogold for light microscopy (LM) or in blots or gels. This chapter includes the following protocols: Protocol for HQ silver enhancement of Nanogold; Protocols for use of silver-enhanced Nanogold with osmium tetroxide--(A) Procedure using reduced concentration of OsO{sub 4}; (B) Procedures for gold toning; Protocol for HQ silver enhancement of Nanogold in pre-embedding immunocytochemistry for cell cultures; Protocol for gold enhancement of Nanogold for EM; Protocol for gold enhancement of Nanogold for LM; Protocol for staining blots with Nanogold and silver enhancement; and Protocol for staining gels with Nanogold and silver enhancement.
Date: April 17, 2002
Creator: Hainfeld, J. F. & Powell, R. D.
System: The UNT Digital Library
Combined Fluorescent and Gold Probes for Microscopic and Morphological Investigations. (open access)

Combined Fluorescent and Gold Probes for Microscopic and Morphological Investigations.

Nanogold{reg_sign}, a gold cluster with a core of gold atoms 1.4 nm in diameter, has proven to be a superior probe label for electron microscopy (EM), giving both higher labeling density and improved access to previously hindered or restricted antigens. It may be visualized by autometallography (AMG) for use in light microscopy (LM): silver-and gold-amplified Nanogold detection has proven to be one of the most sensitive methods available for the detection of low copy number targets such as viral DNA in cells and tissue specimens. AMG enhancement has also made Nanogold an effective detection label in blots and gels. The following protocols will be described: Labeling of nuclear components in cells. Protocol for in situ hybridization and detection with fluorescein-Nanogold--or Cy3{trademark}-Nanogold-labeled streptavidin. Nanogold is an inert molecule, and generally does not interact with biological molecules unless a specific chemical reactivity is introduced into the molecule. Conjugates are prepared using site-specific chemical conjugation through reactive chemical functionalities introduced during Nanogold preparation, which allows the gold label to be attached to a specific site on the conjugate biomolecule. For example, a maleimido-Nanogold derivative, which is specific for thiol binding, is frequently attached to the hinge region of an antibody at a unique …
Date: April 17, 2002
Creator: Powell, R. D. & Hainfeld, J. F.
System: The UNT Digital Library
Texas State Office of Risk Management Strategic Plan: Fiscal Years 2003-2007 (open access)

Texas State Office of Risk Management Strategic Plan: Fiscal Years 2003-2007

Agency strategic plan for the Texas State Office of Risk Management describing the organization's planned services, activities, and other goals during fiscal years 2003 through 2007.
Date: June 17, 2002
Creator: Texas. State Office of Risk Management.
System: The Portal to Texas History
Texas Lottery Commission Strategic Plan: Fiscal Years 2003-2007 (open access)

Texas Lottery Commission Strategic Plan: Fiscal Years 2003-2007

Agency strategic plan for the Texas Lottery Commission describing the organization's planned services, activities, and other goals during fiscal years 2003 through 2007.
Date: June 17, 2002
Creator: Texas Lottery Commission
System: The Portal to Texas History