Colorimetric immunostaining

NDT401a

Tissue preparation & colorimetric immunostaining with 1 primary antibody on free-floating sections (NDT401a-a):

This service includes tissue preparation, sectioning, immunostaining, mounting, coverslipping and labeling the slides. As a result, you will receive up to 60 immunostained 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 cryopretection solution (NDT301). Subsequently, sections cut from various levels (or the levels of your choice) will be processed free-floating for immunohistochemistry with one specific antibody according to the avidin-biotin-complex (ABC) method¹ (cf. see samples below).

Heat shock protein-immunoreactivity. This 30 µm cryostat section was cut from the dorsal hippocampus of a rat that survived for 15 hrs after the injection of aminooxyacetic acid into the entorhinal cortex (for details, cf. Neurosci. Lett. 147:185-188, 1992). The section was processed free-floating according to avidin-biotin-complex method.

Heat shock protein-immunostained section counterstained with thionin. This 30 µm cryostat section was cut from the dorsal hippocampus of a rat that survived for 15 hrs after the injection of aminooxyacetic acid into the entorhinal cortex (for details, cf. Neurosci. Lett. 147:185-188, 1992). The section was processed for heat shock protein-immunostaining (brown) and then counterstained with thionin.

OX18-immunostained section counterstained with thionin. This 30 µm cryostat section was cut from the entorhinal cortex of a rat that survived for 24 hrs after the injection of aminooxyacetic acid into the entorhinal cortex. The section was processed for OX-immunostaining and then counterstained with thionin. Note the expression of OX-18 immunoreactivity (brown) in activated microglia within layer III.

GFAP-immunostained section counterstained with thionin. This 30 µm cryostat section was cut from the entorhinal cortex of a normal rat. The section was processed for GFAP-immunostaining and then counterstained with FD thionin solution (cf. Products, Cat. #PS101). Note GFAP-immunoreactivity (brown) in the processes of astrocytes.

Parvalbumin-immunostained section counterstained with thionin. 30 µm cryostat section through the medial entorhinal cortex of a rat that survived for 24 hrs after kainic acid administration, showing the preferential loss of neurons in layer III and relative resistance of parvalbumin neurons (for details, cf. J. Neurosci. 15:6301-6313, 1995).

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. 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 & colorimetric immunostaining with 1 primary antibody on sections mounted on slides (NDT401a-b):

This service includes tissue preparation, sectioning, immunostaining, mounting, coverslipping and labeling the slides. As a result, you will receive up to 60 immunostained sections per brain or per tissue block ready for microscopic observations.

Procedure: Following cryopretection, 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 slides (Slides available upon request). Sections cut from various levels (or the levels of your choice) will be processed on slides for immunostaining with 1 specific antibody according to the avidin-biotin-complex (ABC) method¹ (cf. see samples below).

Cytokeratin 18-immunostained section counterstained with cresyl violet. This 12 µm frozen section of the rat prostate was processed for Cytokeratin 18-immunoreactivity (brown) and was then counterstained with FD cresyl violet solution (cf. Products, Cat. #NDT202).

CD31-immunostained section counterstained with hematoxylin. This 7 µm paraffin section of the mouse ear was processed for CD31-immunoreactivity (brown) and was then counterstained with FD hematoxylin solution (cf. Products, Cat. #NDT204).

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. 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 & colorimetric immunostaining with 2 primary antibodies on free-floating sections (NDT401a-c):

This service includes tissue preparation, sectioning, immunostaining, mounting, coverslipping and labeling of slides. As a result, you will receive up to 60 immunostained 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 immunostaining with 2 specific antibodies according to the avidin-biotin-complex (ABC) method¹ (cf. see samples below).

BrdUand NeuN double-immunostaining. 30 £gm cryostat section was cut from the hippocampal dentate gyrus of a mouse sacrificed 24 hr after the injection with 5-bromo-2-deoxyuridine (BrdU). The section was processed free-floating for BrdU-(blue) and then for NeuN-(red) immunoreactivity according to the ABC-AP method (cf. also photo samples of NDT401b).

C-fos and kir2.3 double immunostaining. 30 £gm cryostat section through the rat hypothalamus was processed free-floating for kir2.3- (brown) and then for c-fos- (red) immunoreactivities according to the avidin-biotin-complex method. Note nuclear labeling of c-fos immunoreactivity.

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. 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 & colorimetric immunostaining with 2 primary antibodies on sections mounted on slides (NDT401a-d):

This service includes tissue preparation, sectioning, immunostaining, mounting, coverslipping and labeling the slide. As a result, you will receive up to 60 immunostained sections per brain or per tissue block ready for microscopic observations.

Procedure: Following cryopretection, 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 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 the avidin-biotin-complex (ABC) method¹.

Remarks:

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

Reference:

  1. 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.