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ANALYTICAL CHEMISTRY · 2023 · IF 7.4

Label-Free Quantification of DNA Hybridization Kinetics and Affinity Enabled by an Electro-Optical Modulation on Single Nanoparticles

Hao Zhu,Zheng Cai,Zixuan Chen,Yun Chen,Jun-Jie Zhu

Since the formation of a DNA duplex plays a vital role in gene expression and regulation across all kinds of organisms, quantifying the interaction during DNA hybridization is essential for understanding various biological processes in living systems on the molecular scale of nucleic acids. Here, we developed a label-free method to measure the binding kinetics and affinity of DNA hybridization with an electro-optical modulation on the individual gold nanorods based on a total internal reflection dark-field imaging microscopy. Under the electrochemical modulation, a Fourier transform filter was utilized to extract the optical scattering of the nanorods, which varies with the DNA binding due to their impedance change. We validated the imaging principle and established an analytical model to monitor the optical response during the DNA hybridization. Using the presented platform, we measured the binding kinetics and affinities of different DNA pairs and demonstrated its capability to distinguish the DNA hybridization with only single-base mismatch, which may provide guidance to explore the etiology and pathogenesis of diseases associated with single point gene mutations. Furthermore, the method allows for simultaneous imaging of multiple nanoparticles, thus enabling a high-throughput detection and opening possibilities for the study of the interfacial heterogeneity.

研究领域: 生物物理学 · 分子生物学 · 纳米技术 · 遗传学
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ANALYTICAL CHEMISTRY · 2021 · IF 6.986

Artificial Intelligence-Aided Multiple Tumor Detection Method Based on Immunohistochemistry-Enhanced Dark-Field Imaging

Lin Fan,Ting Huang,Doudou Lou,Zengzhou Peng,Yongqi He,Xinyu Zhang,Ning Gu,Yu Zhang

The immunohistochemical method serves as one of the most practical tools in clinical cancer detection and thus has great application value to overcome the existing limits of the conventional method and further improve the detecting efficiency and sensitivity. This study employed 3,3′-diaminobenzidine (DAB), a conventional color indicator for immunohistochemistry, as a novel high-sensitive scattering reagent to provide a multidimensional image signal varying with the overexpression rate of tumor markers. Based on the scattering properties of DAB aggregates, an efficient and robust artificial intelligence-aided immunohistochemical method based on dark-field imaging has been established, with improvement in both the imaging quality and interpretation efficiency in comparison with the conventional manual-operated immunohistochemical method. Referencing the diagnosis from three independent pathologists, this method succeeded in detecting HER2 overexpressed breast tumors with a sensitivity of 95.2% and a specificity of 100.0%; meanwhile, it was found to be applicable for non-small-cell lung tumors and malignant lymphoma as well. As demonstrated, this study provided an effective and reliable means for making diagnostic suggestions, which exhibited great potential in multiple tumor pathological detection at low cost.

CAS / MDL: 7722-84-1 9003-99-0
研究领域: 病理学 · 医学影像学 · 生物医学工程 · 肿瘤学
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Nanoscale · 2015 ·Vol. 8 ·期 16 ·8553-8558 · IF 7.394

Enzyme catalysis enhanced dark-field imaging as a novel immunohistochemical method

Lin Fan,Yanyan Tian,Rong Yin,Doudou Lou,Xizhi Zhang,Meng Wang,Ming Ma,Shouhua Luo,Suyi Li,Ning Gu,Yu Zhang

Conventional immunohistochemistry is limited to subjective judgment based on human experience and thus it is clinically required to develop a quantitative immunohistochemical detection. 3,3′-Diaminobenzidin (DAB) aggregates, a type of staining product formed by conventional immunohistochemistry, were found to have a special optical property of dark-field imaging for the first time, and the mechanism was explored. On this basis, a novel immunohistochemical method based on dark-field imaging for detecting HER2 overexpressed in breast cancer was established, and the quantitative analysis standard and relevant software for measuring the scattering intensity was developed. In order to achieve a more sensitive detection, the HRP (horseradish peroxidase)-labeled secondary antibodies conjugated gold nanoparticles were constructed as nanoprobes to load more HRP enzymes, resulting in an enhanced DAB deposition as a dark-field label. Simultaneously, gold nanoparticles also act as a synergistically enhanced agent due to their mimicry of enzyme catalysis and dark-field scattering properties.

CAS / MDL: 16961-25-4
研究领域: 医学影像 · 癌症研究 · 生物化学 · 分子生物学
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COLLOIDS AND SURFACES B-BIOINTERFACES · 2022 · IF 5.999

GMT8 aptamer conjugated PEGylated Ag@Au core-shell nanoparticles as a novel radiosensitizer for targeted radiotherapy of glioma

Dongdong Li,Jing Zhao,Jun Ma,Huiquan Yang,Xiaodong Zhang,Yuyu Cao,Peidang Liu

Radiotherapy is one of the main treatment modalities for glioma, but the therapeutic efficacy is often limited by the radioresistance of tumor cells. The radiosensitization effects of silver nanoparticles (AgNPs) and gold nanoparticles (AuNPs) on tumors have been confirmed by previous studies. To enhance the specific killing effect of irradiation on tumor cells, targeted modification of radiosensitizers is urgently needed. Herein, we developed polyethylene glycol (PEG)-coated [email protected] core-shell nanoparticles (PSGNPs) modified with GMT8 aptamer (GSGNPs) and evaluated their radiosensitization effects on glioma cells through in vivo and in vitro experiments. Transmission electron microscope image showed that the prepared PSGNPs had a spherical core-shell structure with an average size of 11 nm. The ultraviolet-visible absorption spectra and Fourier transform infrared spectra displayed that GMT8 was successfully conjugated to PSGNPs. The results of dark-field imaging revealed that the targeting ability of GSGNPs to U87 glioma cells was much better than that to normal human microvascular endothelial cells. Additionally, it was also found that the endocytic pathways of GSGNPs mainly involved clathrin-mediated endocytosis and macropinocytosis . The sensitization enhancement ratio of GSGNPs was calculated to be 1.62, which was higher than that of PSGNPs. In vivo imaging results showed that GSGNPs exhibited good tumor targeting and retention capabilities, and the fluorescence intensity ratio of Cy5-GSGNPs to Cy5-PSGNPs reached a peak at 4 h after injection. More importantly, the median survival time of mice bearing U87 glioma was significantly prolonged after intravenous administration of GSGNPs combined with radiotherapy. This work demonstrated that GSGNPs could be used as an effective nano-radiosensitizer for targeted radiotherapy of glioma.

研究领域: 癌症治疗 · 纳米技术 · 放射生物学 · 药学
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