Background: Immunologic heterogeneity is known to exist within both eczematous dermatitis and psoriasis; however, selection of molecularly targeted therapies for individual patients generally does assess for or incorporate such information about patient-specific immune changes. Objective: We sought to develop a rapid, non-invasive method for obtaining and analyzing epidermal protein biomarkers from skin and utilize this methodology to dissect immunologic heterogeneity in both psoriasis and eczematous dermatitis. Methods: We optimize and evaluate detergent-based immune profiling system (DIPS) which utilizes a combination of two detergents to solubilize full-thickness epidermis when applied to the skin with an applicator. Downstream proteomic profiling of this material allows high-throughput immunologic characterization of immune biomarkers. Results: DIPS was performed on 43 patients with psoriasis and 27 patients with eczematous dermatitis. This approach was found to be painless, nonscarring, and enabled rapid turnaround from sample collection to data output. We used this approach to accurately differentiate psoriasis and eczema using a limited set of proteins and to identify cases of eczema/psoriasis overlap with non-canonical molecular profiles. Additionally, we measured patient-specific cytokine profiles in eczema that correlated with response to IL-4/IL-13 blockade. Conclusion: DIPS is a promising new non-invasive cutaneous immune profiling approach that can deconvolute immune heterogeneity amongst patients with both psoriasis and eczematous dermatitis.