NeuroScience Associates

Neurohistology Stains

MultiBrain® Technology provides a tremendous foundation from which NSA is able to provide efficient, low cost services. Complementing this enabling technology is our staining service where clients send their tissue to NSA for embedding and sectioning, followed by the application of their desired stain(s). Specifically, NSA Labs® strives to offer a wide range of stains including traditional/classic, immunohistochemistry and a variety of proprietary and specialty stains specifically designed to reveal unique features in tissues.

NSA Labs® emphasizes high contrast staining results as a standard by which all of our stains are measured (traditional or proprietary).

Below are common traditional/classic stains requested of NSA. If you don’t see a stain you desire, contact NSA. Also, visit our Antibody Gallery for further information on our antibody work.

Traditional/Classic Stains

Thionine: To reveal cell bodies in tissue, may be applied as a counterstain against suitable stains

H&E: To reveal the nuclei and cytoplasm of cell bodies

Weil-Myelin: To reveal the myelinated axons

Solochrome: To reveal myelin

Perls (with or without DAB): To detect the presence of ferric iron in tissue (normally occurring, or from ruptured red blood cells)

Fluoro-Jade: To reveal degenerating neurons

Congo Red: To reveal fibrillar amyloid plaques

Thioflavine S: To reveal fibrillar amyloid

Specialty Stains

Silver-Based Stains

Silver, which has long been used for its black color and development capabilities in black and white film, is successfully utilized by NSALabs® in many of the specialty stains. Specially devised preparation of tissues creates an affinity for silver to bind to unique features in brain sections. Different preparations are used to direct silver to bind to completely different structures, allowing for the creation of unique stains. When the silver is developed it becomes pure black which creates the ultimate contrast against unstained tissue. Directing the silver to the appropriate tissue components is an art that NSA has perfected for revealing a variety of unique characteristics in some of our specialty stains.

Amino Cupric Silver (Amino CuAg) /Neutral Red Counter: To reveal degeneration

Cupric Silver (CuAg): To reveal degeneration in fragile tissue

Campbell-Switzer Alzheimer: To reveal amyloid plaques and tau abnormalities

Autometallography: To reveal metals

Blood Brain Barrier Compromise (BBB)

BBB compromise can be revealed with immunohistochemistry by using an antibody against the animal’s own IgG. Areas with “leaky” BBB allow serum proteins to pass into the brain parenchyma, e.g., IgG’s and albumin.

Blood Brain Barrier (BBB): To reveal the locations of blood brain barrier compromise in the brain

Ischemia Contrast

This staining method delineates the penumbra while highlighting the infarcted volume in sharp contrast. Normal tissue is dense black, infarcted areas fail to stain and gradations of gray identify the penumbra zone. Section images are digitized and, based on optical density, the volumes of the infarct, penumbra and normal brain are calculated. Adjacent sections stained with other methods including H&E, thionine and GFAP IHC display less definition of affected areas.

Ischemia Contrast: To reveal the volume of tissue affected following ischemia

Immunohistochemistry/Antibody Stains

Immunohistochemistry (IHC) is used to detect the presence of specific biomolecules (antigens) in tissue sections by applying an antibody to that antigen. IHC has become one of the most common tools in neurohistologic research due to the potential of high specificity when staining a target feature. NSA uses TritonX-100 to fenestrate membranes, allowing antibodies to stain the entire thickness of the section (30–80µ), removing thickness as a factor in NSA’s protocols.

We offer high-quality immunohistochemistry services utilizing commercially available antibodies and client-supplied proprietary antibodies and routinely perform titration series to expand our repertoire.

The following images are some of our most commonly requested immunohistochemistry. Complete listing of commercial antibodies that NSA has successfully applied.