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NDT401cc
NDT401-cc Tissue preparation & immunogold labeling with 2 primary antibodies on free-floating sections

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. To express our appreciation, a set of Nissl (or H & E) stained sections adjacent to those used for immunolabeling 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, 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 (photo samples below).

Bcl2 & parvalbumin double immunostaining. 30-micron 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-micron 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-micron cryostat section of the rat cortex was processed free-floating for parvalbumin-immuoreactivity (black deposits) with the immunogold labeling technique and then for GABA-immuoreactivity (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:
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. 

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

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