BackgroundBiomarkers play a key role in risk assessment, assessing treatment response, and detecting recurrence and the investigation of multiple biomarkers may also prove useful in accurate prediction and prognosis of cancers. Immunohistochemistry (IHC) has been a major diagnostic tool to identify therapeutic biomarkers and to subclassify breast cancer patients. However, there is no suitable IHC platform for multiplex assay toward personalized cancer therapy. Here, we report a microfluidics-based multiplexed IHC (MMIHC) platform that significantly improves IHC performance in reduction of time and tissue consumption, quantification, consistency, sensitivity, specificity and cost-effectiveness.Methodology/Principal FindingsBy creating a simple and robust interface between the device and human breast tissue samples, we not only applied conventional thin-section tissues into on-chip without any additional modification process, but also attained perfect fluid control for various solutions, without any leakage, bubble formation, or cross-contamination. Four biomarkers, estrogen receptor (ER), human epidermal growth factor receptor 2 (HER2), progesterone receptor (PR) and Ki-67, were examined simultaneously on breast cancer cells and human breast cancer tissues. The MMIHC method improved immunoreaction, reducing time and reagent consumption. Moreover, it showed the availability of semi-quantitative analysis by comparing Western blot. Concordance study proved strong consensus between conventional whole-section analysis and MMIHC (n = 105, lowest Kendall's coefficient of concordance, 0.90). To demonstrate the suitability of MMIHC for scarce samples, it was also applied successfully to tissues from needle biopsies.Conclusions/SignificanceThe microfluidic system, for the first time, was successfully applied to human clinical tissue samples and histopathological diagnosis was realized for breast cancers. Our results showing substantial agreement indicate that several cancer-related proteins can be simultaneously investigated on a single tumor section, giving clear advantages and technical advances over standard immunohistochemical method. This novel concept will enable histopathological diagnosis using numerous specific biomarkers at a time even for small-sized specimens, thus facilitating the individualization of cancer therapy.

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A microfabricated cell-based testing device for electrochemotherapy (ECT) has been developed; it mimics a clinical electroporator with a circular needle array and maintains a similar electric field strength distribution. Until now, the performance between electroporators having two- and six-needle circular array electrodes, which are the general needle-type clinical electroporators for ECT, has not been evaluated systemically, although many studies have investigated the efficacy of ECT on cancer cells. In this study, the cell-based performance on the newly developed ECT testing device was analyzed in two- and six-electrode modes using propidium iodide and bleomycin, and the electroporation characteristics were characterized. With on-chip fluorescence-based assays, the proposed device offers time- and cost-effective experimental procedures compared with conventional assays, and the results can reflect the electric characteristics of clinical electroporators.

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Anaphylaxis is an acute, potentially lethal, multisystem syndrome resulting from the sudden release of mast-cell- and basophile-derived mediators into the circulation. Common manifestations of anaphylactic reactions include urticaria, angioedema, nausea, vomiting, hypotension and cardiovascular collapse. Cardiovascular collapse is the first detected manifestation in up to 50% of cases in perioperative anaphylaxis, because patients are anesthetized and unable to report symptoms. A 25-year-old male presented with severe hypotension and erythema after intravenous atropine administration during general anesthesia. Postoperative laboratory findings demonstrated elevated serum tryptase and total immunoglobulin E. An intradermal test showed atropine sensitivity. Although atropine is used widely as a perioperative anticholinergic agent, it is a potential risk factor for a severe anaphylactic reaction. Therefore, prompt recognition and adequate therapeutic measures are necessary to avoid fatal consequences.