TY - JOUR
T1 - Automated direct immunofluorescence analyses of skin biopsies
AU - Lemcke, Susanne
AU - Sokolowski, Siegmar
AU - Rieckhoff, Nicole
AU - Buschtez, Michael
AU - Kaffka, Christian
AU - Winter-Keil, Antje
AU - Schaller, Christine
AU - Rottmann, Norbert
AU - Sadik, Christian D.
AU - Stöcker, Winfried
AU - Zillikens, Detlef
AU - Schmidt, Enno
PY - 2016/3/1
Y1 - 2016/3/1
N2 - Background Diagnosis of autoantibody- and immune complex-induced skin diseases is primarily based on direct immunofluorescence (DIF) microscopy. DIF staining is usually performed manually and, therefore, is labor intensive. The quality of immunohistochemical results considerably depends on the experience of the person conducting the tests. The novel EUROTide™ technique in combination with the biochip-based system EUROPath represents a new technology for automation of DIF staining. Methods Frozen sections of previously characterized skin biopsies from bullous pemphigoid and pemphigus vulgaris patients were incubated with fluorescein-labelled anti-human IgG and complement C3c following the standard manual procedure and, for comparison, applying EUROTide/EUROPath in an automated version. Results Both, the manual and the automated procedure, detected IgG and C3c deposits in all samples. However, DIF stainings performed with EUROTide/EUROPath displayed more intense specific IF signals and distinctly less background staining. The detecting antibody could be used at a ×4 higher dilution. Conclusion EUROTide/EUROPath applied in an automated system improves diagnostic accuracy and saves reagents. Larger studies in other routine laboratories may further explore the value of the EUROTide/EUROPath technology and may include comparison with other automated stainers.
AB - Background Diagnosis of autoantibody- and immune complex-induced skin diseases is primarily based on direct immunofluorescence (DIF) microscopy. DIF staining is usually performed manually and, therefore, is labor intensive. The quality of immunohistochemical results considerably depends on the experience of the person conducting the tests. The novel EUROTide™ technique in combination with the biochip-based system EUROPath represents a new technology for automation of DIF staining. Methods Frozen sections of previously characterized skin biopsies from bullous pemphigoid and pemphigus vulgaris patients were incubated with fluorescein-labelled anti-human IgG and complement C3c following the standard manual procedure and, for comparison, applying EUROTide/EUROPath in an automated version. Results Both, the manual and the automated procedure, detected IgG and C3c deposits in all samples. However, DIF stainings performed with EUROTide/EUROPath displayed more intense specific IF signals and distinctly less background staining. The detecting antibody could be used at a ×4 higher dilution. Conclusion EUROTide/EUROPath applied in an automated system improves diagnostic accuracy and saves reagents. Larger studies in other routine laboratories may further explore the value of the EUROTide/EUROPath technology and may include comparison with other automated stainers.
UR - http://www.scopus.com/inward/record.url?scp=84946426666&partnerID=8YFLogxK
U2 - 10.1111/cup.12637
DO - 10.1111/cup.12637
M3 - Journal articles
C2 - 26454250
AN - SCOPUS:84946426666
SN - 0303-6987
VL - 43
SP - 227
EP - 235
JO - Journal of Cutaneous Pathology
JF - Journal of Cutaneous Pathology
IS - 3
ER -