Tissue preparation & immunofluorescence labeling with 1 primary antibody on free-floating sections

Cat. # SI201  

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. #PC101). 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 hr 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.