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Single virus fingerprinting by widefield interferometric defocus-enhanced mid-infrared photothermal microscopy

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Version 2 2023-08-09, 16:00
Version 1 2023-01-10, 03:34
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posted on 2023-08-09, 16:00 authored by Qing Xia, Zhongyue Guo, Haonan Zong, Scott Seitz, Celalettin Yurdakul, M. Selim Unlu, Le Wang, John H. Connor, Ji-Xin Cheng
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 acids signatures of thymine and uracil residue vibrations are obtained to differentiate DNA and RNA viruses. WIDE-MIP imaging further reveals an enriched \b{eta} 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.

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