Immunofluorescence Protocol Frozen Section

Cut 4 8 um thick cryostat sections and mount on superfrost plus slides or gelatin coated slides.

Immunofluorescence protocol frozen section.

Tissue preparation perfusion and fixation note. Dry the tissue sections overnight at room temperature. This protocol is also suitable for 40µm free floating. This ihc protocol provides a basic guide for the fixation cryostat sectioning and staining of frozen tissue samples.

Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining. Immunofluorescence on frozen tissue sections bio protocol. 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. For fresh unfixed frozen tissue fix immediately as follows.

Nagy gertsenstein vintersten and behringer ed. Store slides at 80 ºc until needed. The fluorescent immunohistochemistry immunofluorescence protocol below is intended for the fluorescent visualization of protein expression in frozen tissue sections. Place the tissue sections onto glass slides suitable for immunohistochemistry e g.

Store frozen blocks at 80 ºc. Immunocytochemistry and immunofluorescence protocol related fluorescence. Sections can be stored in a sealed slide box at 80 c for later use. Section the frozen tissue block into a desired thickness typically 5 10 µm using the cryotome.

Immunofluorescence can also be used as a qualitative measure of protein expression. This portion of the protocol can be skipped if you are working with pre mounted tissue slides. Microscope slides pre coated. Brigitte arduini version 1 2015 mar 23.

Materials phosphate buffered saline pbs 1x paraformaldehyde pfa 4 see support protocol 1. 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. Cover sections with 4 formaldehyde diluted in warm 1x pbs.

For fixed frozen tissue proceed with immunostaining section c. Icc and if video protocol. Immunofluorescence on frozen sections. Snap frozen fresh tissues in liquid nitrogen or isopentane pre cooled in liquid nitrogen embedded in oct compound in cryomolds.

Protocol for immunofluorescent staining of mouse frozen sections tissue. Modified from manipulating the mouse embryo 3. Direct vs indirect if. Annexin v labeled with alexa fluor 488 in frozen rat placenta section by ihc immunohistochemistry.