Immunofluorescence Protocol Frozen Section

Store slides at 80 ºc until needed.

Immunofluorescence protocol frozen section.

Modified from manipulating the mouse embryo 3. Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds. Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry. Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1.

The following immunohistochemistry ihc protocol has been developed and optimized by r d systems ihc icc laboratory for fluorescent ihc experiments using frozen tissue samples. Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method. Immunofluorescence on frozen sections. Place the tissue sections onto glass slides suitable for immunohistochemistry e g.

Tissue preparation perfusion and fixation note. Nagy gertsenstein vintersten and behringer ed. Sections can be stored in a sealed slide box at 80 c for later use. Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining.

Dry the tissue sections overnight at room temperature. Protocol for immunofluorescent staining of mouse frozen sections tissue. Allow sections to fix for 15 min at room temperature. Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest.

Brigitte arduini version 1 2015 mar 23. Immunofluorescence can also be used as a qualitative measure of protein expression. For fresh unfixed frozen tissue fix immediately as follows. Immunocytochemistry and immunofluorescence protocol related fluorescence.

Icc and if video protocol. Store frozen blocks at 80 ºc. Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or gelatin coated slides. The fluorescent immunohistochemistry immunofluorescence protocol below is intended for the fluorescent visualization of protein expression in frozen tissue sections.

Section the frozen tissue block into a desired thickness typically 5 10 µm using the cryotome. Immunofluorescence on frozen tissue sections bio protocol. For fixed frozen tissue proceed with immunostaining section c. 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. Microscope slides pre coated. This ihc protocol provides a basic guide for the fixation cryostat sectioning and staining of frozen tissue samples. Direct vs indirect if.