Ultrafast, Ultrasensitive Detection and Imaging of Single Cardiac Troponin-T Molecules,ACS Sensors

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Ultrafast, Ultrasensitive Detection and Imaging of Single Cardiac Troponin-T Molecules
ACS Sensors ( IF 8.2 ) Pub Date : 2020-10-30 , DOI: 10.1021/acssensors.0c01790
Pavel N. Melentiev _{1,

2} , Lina V. Son _{3,

4} , Denis S. Kudryavtsev ₃ , Igor E. Kasheverov ₃ , Victor I. Tsetlin ₃ , Rinat O. Esenaliev ₅ , Victor I. Balykin _{1,

2}

Affiliation

The fluorescence-based methods of single-molecule optical detection have opened up unprecedented possibilities for imaging, monitoring, and sensing at a single-molecule level. However, single-molecule detection methods are very slow, making them practically inapplicable. In this paper, we show how to overcome this key limitation using the expanded laser spot, laser excitation in a nonfluorescent spectral window of biomolecules, and more binding fluorescent molecules on a biomolecule that increases the detection volume and the number of collected photons. We demonstrate advantages of the developed approach unreachable by any other technique using detection of single cardiac troponin-T molecules: (i) 1000-fold faster than by known approaches, (ii) real-time imaging of single troponin-T molecules dissolved in human blood serum, (iii) measurement of troponin-T concentration with a clinically important sensitivity of about 1 pg/mL. The developed approach can be used for ultrafast, ultrasensitive detection, monitoring, and real-time imaging of other biomolecules as well as of larger objects including pathogenic viruses and bacteria.

中文翻译：

单心脏肌钙蛋白-T分子的超快，超灵敏检测和成像

基于荧光的单分子光学检测方法为单分子水平的成像，监测和传感打开了前所未有的可能性。但是，单分子检测方法非常慢，因此实际上不适用。在本文中，我们展示了如何使用扩展的激光点，在生物分子的非荧光光谱窗口中进行激光激发以及在生物分子上具有更多结合力的荧光分子来克服这一关键限制，从而增加检测量和收集的光子数量。我们展示了开发的方法的优势，该优势是使用检测单个心脏肌钙蛋白-T分子的任何其他技术无法实现的：（i）比已知方法快1000倍，（ii）实时溶解在人体内的肌钙蛋白-T分子的实时成像血清（iii）测定肌钙蛋白T的浓度，临床上重要的敏感性约为1 pg / mL。所开发的方法可用于其他生物分子以及包括病原病毒和细菌在内的较大物体的超快，超灵敏检测，监视和实时成像。

更新日期：2020-11-25

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