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NDT401-bc
NDT401-bc Tissue preparation & immunofluorescence labeling with 2 primary antibodies on free-floating sections

This service includes tissue preparation, sectioning, immunostaining, mounting, coverslipping and labeling the slides. As a result, you will receive up to 60 immunostained per brain or per tissue block ready for microscopic observations. To express our appreciation, a set of Nissl (or H & E) stained sections adjacent to those used for immunostaining will also be provided for you without additional cost.

Procedure: Following cryoprotection, tissue will be rapidly frozen in isopentane pre-cooled to -70ºC. The frozen tissue will then be cut on a cryostat and collected in our unique section cryoprotection solution (cf. Product NDT301). 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. photo samples below).

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

Middel: Confocal 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:
1. A quotation is required before placing an order.
2. The investigator needs to provide unfixed frozen tissue.
3. 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.

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