Immunofluorescence Protocol For Frozen Sections

Brigitte arduini version 1 2015 mar 23.

Immunofluorescence protocol for frozen sections.

Tissue preparation cyropreservation. 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. Modified from manipulating the mouse embryo 3.

Immunofluorescence general protocol important. Immunofluorescence on frozen tissue sections bio protocol. See cryoprotection and processing of embryonic tissue protocol. 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.

This protocol is also suitable for 40µm free floating. Immunohistochemistry protocol for frozen sections. Immunofluorescence on frozen sections. Mount tissue sections onto gelatin or poly l lysine coated slides by placing the cold sections onto warm slides.

Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds. Preparation of slides. Immunofluorescence staining protocol. The section will curl if the specimen is too cold.

Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method. Immunocytochemistry and immunofluorescence protocol related fluorescence. The suggested cryostat temperature is between 15 and 23 c. Store frozen blocks at 80 ºc.

Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or. 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. Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest. Direct vs indirect if.

Immunofluorescence can also be used as a qualitative measure of protein expression. Do not allow frozen tissue to thaw before cutting. Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining. Microscope slides pre coated.

Protocol for immunofluorescent staining of mouse frozen sections tissue. Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry. Slides can be safely stored for 6 12 months at 80 c until ready for staining. Nagy gertsenstein vintersten and behringer ed.

Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1.