Paraffin Section Immunofluorescence Protocol

Paraffin immunofluorescence as a salvage technique.

Paraffin section immunofluorescence protocol.

60 minutes paraffin 2. This guide highlights critical steps in the immunofluorescence protocol and demonstrates how protocol changes can affect the final outcome of your experiment. 60 minutes paraffin 1. 60 minutes paraffin 3.

Section the paraffin embedded tissue block in 4 10 μm thickness slides on a microtome and float in a 37 c water bath containing deionized water. Preparation of paraffin sections for immunohistochemistry a. If the sections still have traces of wax an additional immersion of 5 minutes in xylene may be employed. Download immunofluorescent staining of paraffin embedded tissue protocol as a pdf deparaffinization and rehydration tip.

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. Immunohistochemistry protocol for fixing and sectioning paraffin embedded tissues. Float the sections onto clean glass slides and microwave at 65 c for 15 min then the tissue binds to the glass. Transfer the sections onto a superfrost plus slide.

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. Inadequately fixation can make tissues dehydrated during tissue processing and become hard and brittle. Immunofluorescence general protocol important. Picric acid is optional enhances preservation of morphology in some tissues.

For tissue sections embedded in paraffin fluorescent multiplex. Carry out incubations in a humidified chamber to avoid tissue drying out which will lead to non specific binding and high background staining. Immunohistochemistry ihc protocol paraffin section protocol. Antigenic determinants masked by formalin fixation and paraffin embedding often may be exposed by epitope umasking enzymatic digestion or saponin etc.

85 mm na 2 hpo 4 75 mm kh 2 p0 4 4 paraformaldehyde and 14 v v saturated picric acid ph 6 9. Before moving to alcohol grades step make sure to completely deparaffinize the sections. 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. Do not use this pretreatment with frozen sections or cultured cells that are not paraffin embedded.

2 immunofluorescence if combines the use of antibodies with fluorescence imaging techniques to visualize target. Section paraffin blocks at the desired thickness usually 4 5 µm on a microtome and float on a 40 c water bath containing distilled water.