CONGRESS 2023 - Application and Imaging of Direct Immunofluorescence
Direct immunofluorescence (DIF) techniques provide the gold standard diagnostic tests for a number of autoimmune immunobullous diseases, including pemphigus and pemphigoid and have clinical utility in other autoantibody-mediated disorders. They are typically performed on unfixed, frozen skin, mucosal and renal biopsies. For optimal diagnostic value, material for DIF requires careful preparation and a number of factors contribute to the quality of the assessed tissue sections. In particular, weak specific fluorescence intensity and high non-specific background fluorescence can hinder accurate interpretation. This presentation will provide a brief overview of best practice for optimising DIF performance, with a focus on skin and mucosal biopsies. Troubleshooting and helpful tips will be provided covering areas including specimen preparation and the importance of using an appropriate transport medium, rinsing considerations prior to block preparation, factors influencing embedding and cryotomy, protocols for optimising fluorescence and appropriate use of conjugates, imaging considerations for digital pathology solutions, storage of material prior to reporting and best practice strategies for maintaining internal and external quality control, with reference to the NEQAS CPT DIF EQA scheme.
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Learning outcomes
This presentation will provide a brief overview of best practice for optimising performance of direct immunofluorescence on skin and mucosal biopsies, including troubleshooting and helpful tips on specimen preparation, processing, quality control, imaging and storage of specimens and material.