Microscope slides pre coated.
Immunofluorescence protocol for frozen sections.
Preparation of slides.
Slides can be safely stored for 6 12 months at 80 c until ready for staining.
Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or.
Paraffin and frozen sections reagents can be applied manually by pipette or this protocol can be adapted for automated and semi automated systems if these are available.
Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1.
Direct vs indirect if.
See cryoprotection and processing of embryonic tissue protocol.
Store frozen blocks at 80 ºc.
Mount tissue sections onto gelatin or poly l lysine coated slides by placing the cold sections onto warm slides.
Do not allow frozen tissue to thaw before cutting.
Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest.
Immunocytochemistry and immunofluorescence protocol related fluorescence.
Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method.
This portion of the protocol can be skipped if you are working with pre mounted tissue slides.
Immunofluorescence on frozen tissue sections bio protocol.
Nagy gertsenstein vintersten and behringer ed.
The following is a general procedure guide for preparation and staining of acetone fixed frozen tissues using a purified unconjugated primary antibody biotinylated secondary antibody and streptavidin horseradish peroxidase sav hrp and dab detection system.
This protocol is also suitable for 40µm free floating.
Protocol for immunofluorescent staining of mouse frozen sections tissue.
Brigitte arduini version 1 2015 mar 23.
The suggested cryostat temperature is between 15 and 23 c.
The section will curl if the specimen is too cold.
Embed the tissue completely in oct compound prior to cryostat sectioning.
Cut cryostat sections at 5 10 µm and mount on gelatin coated histological slides.
Icc and if video protocol.
Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds.
Immunofluorescence general protocol important.
Modified from manipulating the mouse embryo 3.
Please refer to the applications section on the front page of product datasheet or product webpage to determine if this product is validated and approved for use on cultured cell lines if ic paraffin embedded samples if p or frozen tissue sections if f.
Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry.
Tissue preparation cyropreservation.
Immunofluorescence staining protocol.
Immunohistochemistry protocol for frozen sections.
