Immunofluorescence Protocol For Paraffin Embedded Sections

Float the sections in a 56 c water bath.

Immunofluorescence protocol for paraffin embedded sections.

Thaw mount the sections onto gelatin coated histological slides. Cut 5 15 µm thick tissue sections using a rotary microtome. Ihc protocol video for paraffin embedded tissue sections from cell signaling technology cst cst protocols. Embed the tissue in a paraffin block.

Before moving to alcohol grades step make sure to completely deparaffinize the sections. The section will curl if the specimen is too cold. Slides are pre coated with gelatin to enhance adhesion of the. Embed the tissue in paraffin at 58 c.

The suggested cryostat temperature is between 15 and 23 c. Immunofluorescence general protocol important. The paraffin tissue block can be stored at room temperature for years. Do not use this pretreatment with frozen sections or cultured cells that are not paraffin embedded.

Tissues can be embedded into paraffin using specialized automated tissue processing systems. Rinse sections in pbs tween 20 for 2x2min. Studies performed as early as the 1970s have shown that if p is a valuable salvage technique in renal pathology when frozen tissue is inadequate e g lacks glomeruli or not available 8 10 14 18 20 21 overall diagnostic results by if p can be obtained in 80 of cases 14 18 20 21 but the diagnostic yield varies depending on 3. Transfer the sections onto glass slides suitable for immunohistochemistry e g.

Immunohistochemistry protocol for paraffin embedded tissue sections advertisement doi. Mount the sections onto either probeon or probeon plus or gelatin coated histological slides. A fluorescent dye is a fluorophore that is a molecule that absorbs light energy at a specific wavelength by a process called excitation and then immediately releases the energy at a different wavelength known as emission. Section the paraffin embedded tissue block at 5 8 µm thickness on a microtome and float in a 40 c water bath containing distilled water.

Immunofluorescence microscopy is a method by which a protein can be visualized inside cells using fluorescent dyes. Antigenic determinants masked by formalin fixation and paraffin embedding often may be exposed by epitope umasking enzymatic digestion or saponin etc. April 10 2014 katie crosby 1 jessica simendinger 1 christopher grange 1 michelle ferrante 1 trisha bernier 1 claire standen 1. Cut 5 15 µm thick tissue sections using a cryostat.

If it is too warm it will stick to the knife. If the sections still have traces of wax an additional immersion of 5 minutes in xylene may be employed.