Antibodies are quintessential affinity reagents for the investigation and determination of a protein's expression patterns, localization, quantitation, modifications, purification, and functional understanding. Antibodies are typically used in techniques such as Western blot, immunohistochemistry (IHC), and enzyme-linked immunosorbent assays (ELISA), among others. The methods employed to generate antibodies can have a profound impact on their success in any of these applications. We raised antibodies against 10 serum proteins using 3 immunization methods: peptide antigens (3 per protein), DNA prime/protein fragment-boost (“DNA immunization”; 3 per protein), and full length protein. Antibodies thus generated were systematically evaluated using several different assay technologies (ELISA, IHC, and Western blot). Antibodies raised against peptides worked predominantly in applications where the target protein was denatured (57% success in Western blot, 66% success in immunohistochemistry), although 37% of the antibodies thus generated did not work in any of these applications. In contrast, antibodies produced by DNA immunization performed well against both denatured and native targets with a high level of success: 93% success in Western blots, 100% success in immunohistochemistry, and 79% success in ELISA. Importantly, success in one assay method was not predictive of success in another. Immunization with full length protein consistently yielded the best results; however, this method is not typically available for new targets, due to the difficulty of generating full length protein. We conclude that DNA immunization strategies which are not encumbered by the limitations of efficacy (peptides) or requirements for full length proteins can be quite successful, particularly when multiple constructs for each protein are used.

The delivery of macromolecules into living cells is challenging since in most cases molecules are endocytosed and remain in the endo‐lysosomal pathway where they are degraded before reaching their target. Here, a method is presented to selectively improve cell membrane permeability by nanosecond laser irradiation of gold nanorods (GNRs) with visible or near‐infrared irradiation in order to deliver proteins across the plasma membrane, avoiding the endo lysosomal pathway. GNRs were labeled with the anti‐EGFR (epidermal growth factor receptor) antibody Erbitux to target human ovarian carcinoma cells OVCAR‐3. Irradiation with nanosecond laser pulses at wavelengths of 532 nm or 730 nm is used for transient permeabilization of the cell membranes. As a result of the irradiation, the uptake of an anti‐Ki‐67 antibody was observed in about 50 % of the cells. The results of fluorescence lifetime imaging show that the GNR detached from the membrane after irradiation.

Pathological diagnosis plays an important role in the diagnosis and treatment of hepatocellular carcinoma (HCC). The traditional method of pathological diagnosis of most cancers requires freezing, slicing, hematoxylin and eosin staining, and manual analysis, limiting the speed of the diagnosis process. In this study, we designed a one‐dimensional convolutional neural network to classify the hyperspectral data of HCC sample slices acquired by our hyperspectral imaging system. A weighted loss function was employed to promote the performance of the model. The proposed method allows us to accelerate the diagnosis process of identifying tumor tissues. Our deep learning model achieved good performance on our data set with sensitivity, specificity, and area under receiver operating characteristic curve of 0.871, 0.888, and 0.950, respectively. Meanwhile, our deep learning model outperformed the other machine learning methods assessed on our data set. The proposed method is a potential tool for the label‐free and real‐time pathologic diagnosis. © 2019 International Society for Advancement of Cytometry

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The electrical properties and microstructures of ZnO-doped Nd(Co1/2Ti1/2)O3 thin films prepared by rf-magnetron sputtering on indium tin oxide (ITO)/glass substrates at different rf powers and Ar/O2 ratios have been investigated. X-ray diffraction pattern analysis showed that the deposited films exhibited a polycrystalline microstructure. All films exhibited the ZnO-doped Nd(Co1/2Ti1/2)O3 orientation perpendicular to the substrate surface, and the uniformity of surface morphologies of the film increased with decreasing Ar partial pressure and rf power. At an Ar/O2 ratio of 80/20 and a rf power of 250 W, the ZnO-doped Nd(Co1/2Ti1/2)O3 films possess a dielectric constant of 28.6 at 1 kHz, a dissipation factor of 0.02 at 1 kHz, and a leakage current density of 3.6×10-10 A/cm2 at an electrical field of 20 kV/cm.

In situ epitaxial growth of YBa2Cu3O7−x superconducting thin films on (100)BaF2 substrates has been successful by an off-axis dc planar magnetron sputtering method. The zero-resistance temperatures of these thin films are about 81–84 K with transition width of 1.5 K. The characteristics of the films were examined by x-ray diffraction, reflection high energy electron diffraction, Rutherford backscattering spectroscopy ion channeling techniques, and scanning electron microscopy. The experimental results suggest that BaF2 is a promising material as substrates or buffer layers for the epitaxial growth of high Tc superconducting thin films, especially in high frequency applications. Discussions on the heteroepitaxy of the YBCO/BaF2 system are propounded.

GeO 2 thin films were prepared by sol-gel method on ITO/Glass substrate. The electrical and optical properties and the microstructures of these films were investigated with special emphasis on the effects of an annealing treatment in ambient air. The films were annealed at various temperatures from 500 °C to 700 °C. Structural analysis through X-ray diffraction (XRD) and atomic force microscope (AFM) showed that surface structure and morphological characteristics were sensitive to the treatment conditions. The optical transmittance spectra of the GeO 2 /ITO/Glass were measured using a UV-visible spectrophotometer. All films exhibited GeO 2 (101) orientation perpendicular to the substrate surface where the grain size increased with increasing annealing temperature. The optical transmittance spectroscopy further revealed high transparency (over 70%) in the wave range 400 -800 nm of the visible region. At an annealing temperature level of 700 °C, the GeO 2 films were found to possess a leakage current density of 1.31×10-6 A/cm 2 at an electrical field of 20 kV/cm.