Detection of Neurodegeneration

Tissue preparation & neurodegeneration detection with FD NeuroSilver Kit (NDT402a):

This service includes tissue preparation, sectioning, silver-staining, mounting, coverslipping and labeling of slides. As a result, you will receive up to 60 silver-stained sections per brain or per tissue block ready for microscopic observations.

Procedure: Following cryoprotection, tissue will be rapidly frozen in isopentane pre-cooled to -70°C. The frozen tissue will be cut on a cryostat and collected in phosphate buffer containing 4% paraformaldehady. Subsequently, sections cut through various levels (or the levels of your choice) will be processed free-floating for the detection of neurons undergoing degeneration with FD NeuroSilver™ Kit.

FD NeuroSilver™ Kit (cf. Products, Cat. #NDT103) is designed for the detection of degenerating neurons in fixed tissue sections of the central nervous system from experimental animals. This kit has been used widely in animal studies under various experimental conditions. The kit has been proven extremely specific and sensitive for the detection of degenerating neuronal somata, axons, and terminals, as well as amyloid plaques in both the brain and spinal cord (see samples below). It is particularly useful for the detection of small numbers of degenerating neurons that may not be demonstrable with routine histopathological techniques. In addition, this kit may be used in combination with immunohistochemistry (see samples below).

Remarks:

• A quotation is required before placing an order.
• The investigator needs to provide fixed tissue.
• Please contact us for more information.

Tissue preparation & neurodegeneration detection with Gallyas silver staining technique (NDT402b):

This service includes tissue preparation, sectioning, silver-staining, mounting, coverslipping and labeling of slides. As a result, you will receive up to 40 silver-stained sections per brain or per tissue block ready for microscopic observations.

Procedure: Following cryoprotection, tissue will be rapidly frozen in isopentane pre-cooled to -70°C. The frozen tissue will then be cut on a cryostat and collected in our unique section cryoprotection solution (cf. Products, Cat. #NDT301). Subsequently, sections cut through various levels (or the levels of your choice) will be processed free-floating for the detection of neurons undergoing degeneration with Gallyas silver staining technique¹.

This technique was originally described by Gallyas et al.¹ and later modified². It is particularly useful for the detection of early neuronal injury in the central nervous system of experimental animals. Typically, both the neuronal perikarya and processes are silver-stained. This technique permits the morphological categorization of damaged neurons and the detection of subtle changes in the morphology of cell bodies and processes (cf. see samples below)

Remarks:

• A quotation is required before placing an order.
• The investigator needs to provide tissue fixed by perfusion with a special fixative.
• Please contact us for more information.

References:

1. Gallyas et al. (1990) Acta Neuropath. 79, 620-628.
2. Du et al. (1998) Neuroscience 82, 1165-1178.

Tissue preparation & neurodegeneration detection with in situ nick translation technique (ISNT) (NDT402c):

This service includes sectioning, ISNT labeling, coverslipping and labeling of slides. As a result, you will receive up to 40 ISNT-labeled sections per brain or per tissue block ready for microscopic observations.

Procedure: Unfixed frozen tissue will be cut on a cryostat and mounted on proteinase-resistant microscope slides (Slides available upon request). Sections cut through various levels (or the levels of your choice) will be processed on slides for the detection of neurodegeneration with in situ nick translation technique¹.

This technique, originally described by Gold et al. (1993)¹, labels the nuclei of neurons undergoing DNA fragmentation. This method is based on the ability of DNA polymerase I to catalyze the polymerization of deoxyuridines onto the ends of DNA fragments. The integrated biotins are amplified and visualized by the avidin-biotin-complex methods² (cf. see samples below).

Remarks:

• A quotation is required before placing an order.
• The investigator needs to provide unfixed frozen tissue.
• Please contact us for more information.

References:

1. Gold R., Schmied M., Rothe G., Zischler H., Breitschopf H., Wekerle H. and Lassmann H. (1993) Detection of DNA fragmentation in apoptosis: application of in situ nick translation to cell culture systems and tissue sections. J. Histochem. Cytochem. 41: 1023-1030.
2. Hsu S. M., Raine L. and Fanger H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29: 577-580.

Tissue preparation & neuronal apoptosis detection with in situ DNA nick end-labeling technique (TUNEL) (NDT402d:):

This service includes sectioning, TUNEL labeling, coverslipping and labeling of slides. As a result, you will receive up to 40 TUNEL-labeled sections per brain or per tissue block ready for microscopic observations.

Procedure: Unfixed frozen tissue will be cut on a cryostat and mounted on proteinase-resistant microscope slides (Slides available upon request). Sections cut through various levels (or the levels of your choice) will then be processed on slides for the detection of neurons undergoing apoptosis with in situDNA nick end-labeling¹.

This technique, originally described by Gavrieli et al. (1992)¹, labels the nuclei of neurons undergoing DNA fragmentation. This method is based on the ability of terminal deoxynucleotidyl transferase to catalyze incorporation of biotinylated deoxyuridines onto the free 3'-hydroxyl termini of DNA fragments, which are considered as one of the most characteristic features of apoptosis²^, ³. The integrated biotins are amplified and visualized by the avidin-biotin-complex (ABC) method⁴ (cf. see samples below).

Remarks:

• A quotation is required before placing an order.
• The investigator needs to provide unfixed frozen tissue.
• Please contact us for more information.

References:

1. Gavrieli Y, Sherman Y, and Ben-Sasson SA: Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J. Cell Biol. 119: 493-501, 1992.
2. Wyllie A. H. (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284: 555-556.
3. Arends M. J., Morris R. G. and Wyllie A. H. (1990) Apoptosis: the role of the endonuclease. Amer. J. Pathol. 136: 593-608.
4. Hsu S. M., Raine L. and Fanger H. (1981) Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29: 577-580.

Terms and Conditions

• For quality assurance of our service, it is recommended that you discuss with us for preferred perfusion protocol and histology and/or immunolabeling protocols.
• It is suggested that you use Gel-coated microscopic slides for tissue mounting and 0.17um-thick coverslips.
• A 15% of the fee will be due upon authorization of the study; and the remaining fee will be due upon delivery of study results.
• Progress of the service is contingent upon staining quality of tissues, operated by the independent contractor.
• Should early termination occur, Neurodigitech will prorate the cost incurred and invoice the Sponsor. The first portion of the fee is non-refundable.