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

Cat. # SI303  

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.

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. #PC101). 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 (cf. photo samples below).
                Bcl2 & parvalbumin double immunostaining.  30 µm 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 µm 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 µm cryostat section of the rat cortex was processed free-floating for parvalbumin-immunoreactivity (black deposits) with the immunogold labeling technique and then for GABA-immunoreactivity (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: 

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