Immunofluorescence Labeling

NDT401b

Tissue preparation & immunofluorescence labeling with 1 primary antibody on free-floating sections (NDT401b-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 cryoprotection solution (cf. Products, Cat. #NDT301). Subsequently, sections cut from various levels (or the levels of your choice) will be processed free-floating for immunostaining with 1 specific antibody according to the indirect immunofluorescence method¹ (cf. photo samples below).

Cofocal image of BrdU-immunoreactivity. 30 µm cryostat section was cut from the hippocampal dentate gyrus of a mouse that survived for 24 hrs after the injection with 5-bromo-2-deoxyuridine (BrdU). This section was processed free-floating according to the indirect fluorescence method.

Cofocal image of NeuN-immunoreactivity. The same section as shown above was processed free-floating for NeuN-immuoreactivity according to the indirect fluorescence method. Note NeuN-labeled granule cells and polymorphic neurons in the dentate gyrus.

Colocalization of BrdU- and NeuN- immunoreactivities. A digital overlay of the 2 images shown above. Note that the regions of colocalization, reflecting the additive effect of superimposed green and red pixels, appear in yellow.

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. Coons, A.H. (1958) Fluorescent antibody methods. In J.F. Danielli (ed): General Cytochemical Methods. New York: Academic Press, pp. 399-422.

Tissue preparation & immunofluorescence labeling with 1 primary antibody on sections mounted on slides (NDT401b-b):

This service includes tissue preparation, sectioning, immunostaining, 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 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 one specific antibody according to the indirect immunofluorescence method¹ (cf. photo samples below).

Substance P-immunofluorescence in the spinal cord. 10 µm cryostat section was cut transversely from the chicken spinal cord. This section was processed on slide according to the indirect fluorescence method (for details, cf. J. Comp. Neurol. 278:253-264, 1988). Note substance P containing fibers mainly in the dorsolateral funiculus, Lissauer’s tract and the dorsal horn.

Substance P-immunofluorescence in the spinal cord. A 10 µm cryostat section of the chicken spinal cord was processed as described above. Note dense substance P immunoreactivity in the dorsolateral funiculus and the dorsal horn.

Vasoactive intestinal polypeptide-immunofluo-rescence in the spinal cord. A 10 µm cryostat section of the chicken spinal cord was processed on slide according to the indirect fluorescence method. Note 2 large neurons containing vasoactive intestinal polypeptide in the nucleus of the dorsolateral funiculus (for details, cf. J. Comp. Neurol. 278:253-264, 1988).

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. Coons, A.H. (1958) Fluorescent antibody methods. In J.F. Danielli (ed): General Cytochemical Methods. New York: Academic Press, pp. 399-422.

Tissue preparation & immunofluorescence labeling with 2 primary antibodies on free-floating sections (NDT401b-c):

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 cryoprotection solution (cf. Products, Cat. #NDT301). Subsequently, sections cut from various levels (or the levels of your option) will be processed free-floating for immunostaining with 2 specific antibodies according to the indirect immunofluorescence method¹ (cf. photo samples below).

Left: Cofocal image of BrdU-immunoreactivity. 30 µm cryostat section was cut from the hippocampal dentate gyrus of a mouse that survived for 24 hrs after the injection with 5-bromo-2-deoxyuridine (BrdU). This section was processed free-floating according to the indirect fluorescence method.

Middle: Cofocal image of NeuN-immunoreactivity. The same section as shown on the left was processed free-floating for NeuN-immunoreactivity according to the indirect fluorescence method. Note NeuN-labeled granule cells in the dentate gyrus.

Right: Colocalization of BrdU- and NeuN-immunoreactivities. A digital overlay of the 2 images shown on the left. Note that the regions of colocalization, reflecting the additive effect of superimposed green and red pixels, appear in yellow.

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. Coons, A.H. (1958) Fluorescent antibody methods. In J.F. Danielli (ed): General Cytochemical Methods. New York: Academic Press, pp. 399-422.

Tissue preparation & immunofluorescence labeling with 2 primary antibodies on sections mounted on slides (NDT401b-d):

This service includes tissue preparation, sectioning, immunostaining, 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 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 the indirect immunofluorescence method¹.

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. Coons, A.H. (1958) Fluorescent antibody methods. In J.F. Danielli (ed): General Cytochemical Methods. New York: Academic Press, pp. 399-422.

Tissue preparation & immunofluorescence labeling with 3 primary antibodies on free-floating sections (NDT401b-e):

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 cryoprotection solution (cf. Products, Cat. #NDT301). Subsequently, sections cut from various levels (or the levels of your choice) will be processed free-floating for immunostaining with 3 specific antibodies according to the indirect immunofluorescence method¹.

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. Coons, A.H. (1958) Fluorescent antibody methods. In J.F. Danielli (ed): General Cytochemical Methods. New York: Academic Press, pp. 399-422.

Tissue preparation & immunofluorescence labeling with 3 primary antibodies on sections mounted on slides (NDT401b-f):

This service includes tissue preparation, sectioning, immunostaining, 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 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 3 specific antibodies according to the indirect immunofluorescence method¹.

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. Coons, A.H. (1958) Fluorescent antibody methods. In J.F. Danielli (ed): General Cytochemical Methods. New York: Academic Press, pp. 399-422.

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.