Automatic differentiation of u- and n-serrated patterns in direct immunofluorescence images

Epidermolysis bullosa acquisita (EBA) is a subepidermal autoimmune blistering disease of the skin. Manual u- and n-serrated patterns analysis in direct immunofluorescence (DIF) images is used in medical practice to differentiate EBA from other forms of pemphigoid. The manual analysis of serration patterns in DIF images is very challenging, mainly due to noise and lack of training of the immunofluorescence (IF) microscopists. There are no automatic techniques to distinguish these two types of serration patterns. We propose an algorithm for the automatic recognition of such a disease. We first locate a region where u- and n-serrated patterns are typically found. Then, we apply a bank of B-COSFIRE filters to the identified region of interest in the DIF image in order to detect ridge contours. This is followed by the construction of a normalized histogram of orientations. Finally, we classify an image by using the nearest neighbors algorithm that compares its normalized histogram of orientations with all the images in the dataset. The best results that we achieve on the UMCG publicly available data set is 84.6% correct classification, which is comparable to the results of medical experts

Enhanced diagnostic immunofluorescence using biopsies transported in saline

BACKGROUND: The demonstration of tissue-bound immunoreactants by direct immunofluorescence microscopy (DIF) is a valuable parameter in the diagnosis of various autoimmune and immunecomplex-mediated skin diseases. For preservation of tissue-bound immunoreactants, biopsies are usually fresh-frozen in liquid nitrogen or transported in Michel's fixative. But even optimally preserved tissue specimens are no guarantee for the correct diagnosis by DIF, especially when weak to moderate IgG fluorescence of the epidermal basement membrane zone is involved. In such cases false negative results are easily obtained due to the relatively high dermal "background" fluorescence produced by polyclonal anti-human IgG fluorescein conjugates. METHODS: In the present study we have compared the use of normal saline (0.9% NaCl) with liquid nitrogen and Michel's fixative as transport medium for skin biopsies. From 25 patients with an autoimmune skin disease (pemphigus, pemphigoid, lupus erythematosus and vasculitis) four matched skin biopsies were obtained and transported in either saline for 24 and 48 hours, liquid nitrogen, or Michel's fixative for 48 hours. RESULTS: Direct IF microscopy showed significant reduction of background fluorescence (p < 0.01) and relatively enhanced desired specific (IgG, IgA) staining in biopsies transported in saline. A conclusive or tentative IF diagnosis was reached in 92% after 24 h saline, 83% after 48 h saline, 68% after freezing in liquid nitrogen, and 62% after 48 h Michel's medium (n = 25). CONCLUSIONS: We conclude that transporting biopsies without freezing in normal saline for 24 hours is an adequate and attractive method for routine IF diagnosis in autoimmune and immune complex-mediated dermatoses. The superior results with saline incubation are explained by washing away of IgG background in dermis and epidermis