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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.

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.


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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