Immunofluorescence Protocol For Paraffin Embedded Sections

April 10 2014 katie crosby 1 jessica simendinger 1 christopher grange 1 michelle ferrante 1 trisha bernier 1 claire standen 1.

Immunofluorescence protocol for paraffin embedded sections.

Float the sections in a 56 c water bath. The paraffin tissue block can be stored at room temperature for years. Embed the tissue in paraffin at 58 c. Paraffin immunofluorescence as a salvage technique.

The suggested cryostat temperature is between 15 and 23 c. Cut 5 15 µm thick tissue sections using a rotary microtome. Embed the tissue in a paraffin block. Cut 5 15 µm thick tissue sections using a cryostat.

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. Antigenic determinants masked by formalin fixation and paraffin embedding often may be exposed by epitope umasking enzymatic digestion or saponin etc. If the sections still have traces of wax an additional immersion of 5 minutes in xylene may be employed. Before moving to alcohol grades step make sure to completely deparaffinize the sections.

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. Download immunofluorescent staining of paraffin embedded tissue protocol as a pdf deparaffinization and rehydration tip. Thaw mount the sections onto gelatin coated histological slides. Slides are pre coated with gelatin to enhance adhesion of the.

Immunohistochemistry protocol for paraffin embedded tissue sections advertisement doi. If it is too warm it will stick to the knife. Immunofluorescence microscopy is a method by which a protein can be visualized inside cells using fluorescent dyes. The section will curl if the specimen is too cold.

Mount the sections onto either probeon or probeon plus or gelatin coated histological slides. Tissues can be embedded into paraffin using specialized automated tissue processing systems. Transfer the sections onto glass slides suitable for immunohistochemistry e g. Ihc protocol video for paraffin embedded tissue sections from cell signaling technology cst cst protocols.

Immunofluorescence general protocol important. 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.