用于单颗粒蛋白质/核酸定量的紫外直接吸收显微镜
Ultraviolet direct absorption microscopy for single particle protein/nucleic acid quantification
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中文总结 AI 辅助
针对单颗粒生物分子表征挑战，提出紫外直接吸收显微镜（UV-DAM），结合定制照明与光源，实现快速无标记高分辨率成像，能在单病毒水平区分不同病毒衣壳，推动纳米医学表征发展，适用于多种纳米材料及生物过程研究。
中文摘要 AI 辅助
生物纳米颗粒对下一代纳米治疗至关重要，但单颗粒生物分子表征仍是一项关键挑战。本文提出紫外直接吸收显微镜（UV-DAM）来应对这一挑战。UV-DAM基于定制照明方案用于仅吸收成像，并结合定制深紫外光源，能提供生物分子特异性和单颗粒灵敏度，实现快速、无标记、高分辨率的生化成像。通过这些能力，在单病毒水平实施经典核酸：蛋白质吸收测定，基于双分子特异性吸收指纹区分空病毒衣壳和载有DNA的病毒衣壳。UV-DAM实现了从体相紫外可见光谱法到单颗粒评估的转变，对纳米医学表征至关重要，且适用于多种纳米材料或高时空分辨率及固有分子对比度的生物过程研究。
英文摘要
Bio-nanoparticles are pivotal to next generation nanotherapeutics, but providing single-particle biomolecular characterization remains a crucial challenge. Herein we present ultra-violet direct absorption microscopy (UV-DAM) to tackle this challenge. UV-DAM is based on a tailored illumination scheme for absorption-only imaging, combined with a custom deep UV light source. Combined, they provide biomolecular specificity with single particle sensitivity. As such, UV-DAM provides rapid, label-free, high resolution, biochemical imaging. Enabled by these capabilities, we implement the classic nucleic acid:protein absorption assay at the single virus level where we demonstrate UV-DAM's ability to distinguish empty from DNA-loaded viral capsids based on bimolecularly specific absorption fingerprints. UV-DAM presents the translation from bulk UV-visible spectrometry to single-particle assessment, a crucial advancement for nanomedicine characterization where particle loading efficiencies are often heterogenous. Beyond this, UV-DAM is applicable to a wide range of nanomaterials or investigation of biological process with high spatio-temporal resolution and intrinsic molecular contrast.
