Immunofluorescence Protocol For Paraffin Embedded Sections

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.

Immunofluorescence protocol for paraffin embedded sections.

Immunohistochemistry protocol for paraffin embedded tissue sections advertisement doi. Mount the sections onto either probeon or probeon plus or gelatin coated histological slides. Paraffin immunofluorescence as a salvage technique. Thaw mount the sections onto gelatin coated histological slides.

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. April 10 2014 katie crosby 1 jessica simendinger 1 christopher grange 1 michelle ferrante 1 trisha bernier 1 claire standen 1. Ihc protocol video for paraffin embedded tissue sections from cell signaling technology cst cst protocols. The suggested cryostat temperature is between 15 and 23 c.

If it is too warm it will stick to the knife. 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. Immunofluorescence microscopy is a method by which a protein can be visualized inside cells using fluorescent dyes. Tissues can be embedded into paraffin using specialized automated tissue processing systems.

Cut 5 15 µm thick tissue sections using a rotary microtome. Download immunofluorescent staining of paraffin embedded tissue protocol as a pdf deparaffinization and rehydration tip. Cut 5 15 µm thick tissue sections using a cryostat. Immunofluorescence general protocol important.

Rinse sections in pbs tween 20 for 2x2min. Please refer to the applications section on the front page of product datasheet or product webpage to determine if this product is validated and approved for use on cultured cell lines if ic paraffin embedded samples if p or frozen tissue sections if f. Slides are pre coated with gelatin to enhance adhesion of the. Float the sections in a 56 c water bath.

The paraffin tissue block can be stored at room temperature for years. Before moving to alcohol grades step make sure to completely deparaffinize the sections. Embed the tissue in a paraffin block. Antigenic determinants masked by formalin fixation and paraffin embedding often may be exposed by epitope umasking enzymatic digestion or saponin etc.

The section will curl if the specimen is too cold. Transfer the sections onto glass slides suitable for immunohistochemistry e g.