Immunogold labeling

NDT401c

Tissue preparation & immunogold labeling with 1 primary antibody on free-floating sections (NDT401c-a):

This service includes tissue preparation, sectioning, immunolabeling, mounting, coverslipping and labeling the slides. As a result, you will receive up to 60 immunolabeled 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 (Slides available upon request). Subsequently, the sections cut from various levels (or the levels of your choice) will be processed free-floating for immunostaining with 1 specific antibody according to immunogold labeling technique¹ (cf. see samples below).

Tyrosine hydroxylase-immunoreactivity in the rat substantia nigra. 30 µm cryostat section cut coronally from the midbrain of a normal rat. This section was processed free-floating according to the immunogold labeling technique. Note densely labeled neurons in the compact area of the substantia nigra.

Tyrosine hydroxylase-immunoreactivity in the rat substantia nigra. The high magnification of the substantia nigra as shown above. Note that dense black deposits in the cytoplasm of neurons are actually metallic silver grains, which can also be viewed under a microscope with a darkfield condenser.

Remarks:

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

Reference:

  1. Humbel B. M., Sibon O. C. M., Stierhof Y.-D. and Schwarz H. (1995) Ultra-small gold particles and silver enhancement as a detection system in immunolabeling and In Situ Hybridization experiments. J. Histochem. Cytochem. 43, 735-737.

Tissue preparation & immunogold labeling with 1 primary antibody on sections mounted on slides (NDT401c-b):

This service includes tissue preparation, sectioning, immunolabeling, coverslipping and labeling the slides. As a result, you will receive up to 60 immunolabeled 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 mounted on gelatin-coated microscope slides (Slides available upon request). Subsequently, sections cut from various levels (the levels of your choice) will be processed on slides for immunostaining with 1 specific antibody according to immunogold labeling technique¹.

Remarks:

  • A quotation is required before placing an order.
  • The investigator needs to provide fixed (or unfixed frozen) tissue and the specific antibody.
  • Please contact us for more information.

Reference:

  1. Humbel B. M., Sibon O. C. M., Stierhof Y.-D. and Schwarz H. (1995) Ultra-small gold particles and silver enhancement as a detection system in immunolabeling and In Situ Hybridization experiments. J. Histochem. Cytochem. 43, 735-737.

Tissue preparation & immunogold labeling with 2 primary antibodies on free-floating sections (NDT401c-c):

This service includes tissue preparation, sectioning, immunolabeling, mounting, coverslipping and labeling the slides. As a result, you will receive up to 60 immunolabeled 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 (Slides available upon request). Subsequently, sections cut from various levels (or the levels of your choice) will be processed free-floating for immunostaining with 2 specific antibodies according to immunogold labeling technique¹ (cf. photo samples below).

Bcl2 & parvalbumin double immunostaining. 30 µm cryostat section of the rat cortex was processed free-floating for parvalbumin-immunoreactivity (black deposits) with the immunogold labeling technique and then for bcl2-immunoreactivity according to avidin-biotin-complex method (red).

NeuN & bcl2 double immunostaining. 30 µm cryostat section of the rat cortex was processed free-floating for bcl2-immunoreactivity (black deposits) with the immunogold labeling technique and then for NeuN-immunoreactivity according to avidin-biotin-complex method (red). Note metallic silver grains mainly accumulated in the cytoplasm of bcl2-containing neurons.

GABA & parvalbumin double immunostaining. 30 µm cryostat section of the rat cortex was processed free-floating for parvalbumin-immunoreactivity (black deposits) with the immunogold labeling technique and then for GABA-immunoreactivity (red) according to avidin-biotin-complex method. Note that metallic silver grains are accumulated in both parvalbumin-containing neuronal perikarya and processes (probably axon terminals), many of which surrounds GABA neurons.

Remarks:

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

Reference:

  1. Humbel BM, Sibon OCM, Stierhof YD, and Schwarz H. (1995) Ultra-small gold particles and silver enhancement as a detection system in immunolabeling and In

Tissue preparation & immunogold labeling with 2 primary antibodies on sections mounted on slides (NDT401c-d):

This service includes tissue preparation, sectioning, immunolabeling, coverslipping and labeling the slides. As a result, you will receive up to 60 immunolabeled 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 mounted on gelatin-coated microscope slides (Slides available upon request). Subsequently, sections cut from various levels (or the levels of your choice) will be processed on slides for immunostaining with 2 specific antibodies according to immunogold labeling technique¹.

Remarks:

  • A quotation is required before placing an order.
  • Investigator needs to provide fixed (or unfixed frozen) tissue and the specific antibodies.
  • Please contact us for more information.

Reference:

  1. Humbel B. M., Sibon O. C. M., Stierhof Y.-D. and Schwarz H. (1995) Ultra-small gold particles and silver enhancement as a detection system in immunolabeling and In Situ Hybridization experiments. J. Histochem. Cytochem. 43, 735-737.

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.