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Immunofluorescence Patterns in Selected Dermatoses, Including Blistering Skin Diseases Utilizing Multiple
Ana Maria Abreu-Velez1, Juliana Calle-Isaza2, Michael S Howard3
1Department of Immunodermatology, Georgia Dermatopathology Associates, Atlanta, Georgia, USA.
North American Journal of Medical Sciences
|November 26, 2015
Summary
Using multiple fluorochromes in direct and indirect immunofluorescence (DIF and IIF) improves autoimmune blistering disease diagnosis by allowing simultaneous visualization of multiple antigens and aiding in ruling out background staining.
Area of Science:
- Dermatology
- Immunopathology
Background:
- Autoimmune vesiculobullous disorders are diagnosed using clinical, histologic, and immunopathologic features.
- Direct and indirect immunofluorescence (DIF and IIF) with salt-split skin are key diagnostic tools.
- Traditional immunofluorescence uses a single fluorochrome.
Purpose of the Study:
- To compare DIF and IIF using a single fluorochrome versus multiple fluorochromes.
- To evaluate the diagnostic utility of multi-fluorochrome immunofluorescence in autoimmune skin diseases.
Main Methods:
- Twenty cases of autoimmune skin disease were analyzed.
- Direct and indirect immunofluorescence (DIF and IIF) were performed using fluorescein isothiocyanate (FITC) alone and with multiple fluorochromes.
- DNA counterstaining was optionally used with multiple fluorochromes.
Main Results:
- Multiple fluorochromes enabled simultaneous visualization of reactivity in diverse skin areas.
- This contrasts with the limitations observed when using fluorescein isothiocyanate (FITC) alone.
Conclusions:
- Multiple fluorochromes facilitate simultaneous labeling of multiple antigens within the same tissue section.
- This technique aids in the colocalization of unknown antigens with known molecules.
- It is also effective in excluding non-specific 'background' staining, enhancing diagnostic accuracy.
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