Department of Electrical and Computer Engineering, Boston University, Boston, MA, 02215, USA.
Department of Biomedical Engineering, Boston University, Boston, MA, 02215, USA.
Nat Commun. 2023 Oct 20;14(1):6655. doi: 10.1038/s41467-023-42439-4.
Clinical identification and fundamental study of viruses rely on the detection of viral proteins or viral nucleic acids. Yet, amplification-based and antigen-based methods are not able to provide precise compositional information of individual virions due to small particle size and low-abundance chemical contents (e.g., ~ 5000 proteins in a vesicular stomatitis virus). Here, we report a widefield interferometric defocus-enhanced mid-infrared photothermal (WIDE-MIP) microscope for high-throughput fingerprinting of single viruses. With the identification of feature absorption peaks, WIDE-MIP reveals the contents of viral proteins and nucleic acids in single DNA vaccinia viruses and RNA vesicular stomatitis viruses. Different nucleic acid signatures of thymine and uracil residue vibrations are obtained to differentiate DNA and RNA viruses. WIDE-MIP imaging further reveals an enriched β sheet components in DNA varicella-zoster virus proteins. Together, these advances open a new avenue for compositional analysis of viral vectors and elucidating protein function in an assembled virion.
临床鉴定和基础病毒学研究依赖于病毒蛋白或病毒核酸的检测。然而，基于扩增和基于抗原的方法由于颗粒小且化学含量低（例如，水疱性口炎病毒中约有 5000 种蛋白质），无法提供单个病毒粒子的精确组成信息。在这里，我们报告了一种宽场干涉离焦增强中红外光热（WIDE-MIP）显微镜，用于高通量指纹识别单个病毒。通过识别特征吸收峰，WIDE-MIP 揭示了单个 DNA 痘苗病毒和 RNA 水疱性口炎病毒中的病毒蛋白和核酸含量。通过获得胸腺嘧啶和尿嘧啶残基振动的不同核酸特征来区分 DNA 和 RNA 病毒。WIDE-MIP 成像进一步揭示了 DNA 带状疱疹病毒蛋白中富含的β片层成分。总之，这些进展为病毒载体的组成分析和阐明组装病毒粒子中的蛋白质功能开辟了新途径。
Zotero 插件
