Immunofluorescence Protocol Frozen Section

Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining.

Immunofluorescence protocol frozen section.

Sections can be stored in a sealed slide box at 80 c for later use. Protocol for immunofluorescent staining of mouse frozen sections tissue. Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry. Cover sections with 4 formaldehyde diluted in warm 1x pbs.

Section the frozen tissue block into a desired thickness typically 5 10 µm using the cryotome. Dry the tissue sections overnight at room temperature. Cryosections adhered to slides from blocks embedded in oct using the 2 methylbutane isobutene method. Immunofluorescence can also be used as a qualitative measure of protein expression.

Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds. For fresh unfixed frozen tissue fix immediately as follows. Nagy gertsenstein vintersten and behringer ed. Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1.

Immunofluorescence is commonly used to determine the cellular or tissue localization of a protein of interest. This protocol is also suitable for 40µm free floating. Place the tissue sections onto glass slides suitable for immunohistochemistry e g. Immunofluorescence on frozen tissue sections bio protocol.

Store frozen blocks at 80 ºc. Immunofluorescence on frozen sections. Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or gelatin coated slides. Modified from manipulating the mouse embryo 3.

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. Store slides at 80 ºc until needed. Allow sections to fix for 15 min at room temperature. For fixed frozen tissue proceed with immunostaining section c.

Icc and if video protocol. The fluorescent immunohistochemistry immunofluorescence protocol below is intended for the fluorescent visualization of protein expression in frozen tissue sections. Tissue preparation perfusion and fixation note. Direct vs indirect if.

Microscope slides pre coated. 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. Brigitte arduini version 1 2015 mar 23.