Ultrasensitive cardiac Troponin-T Sensing by a single molecules counting method
Pavel N. Melentiev,†,‡;
Lina V. Son,⊥,#;
Denis S. Kudryavtsev,⊥;
Igor E. Kasheverov,⊥;
Victor I. Tsetlin,⊥;
Rinat O. Esenaliev,§;
Victor I. Balykin†,‡;
†Institute of Spectroscopy RAS, Troitsk, Moscow 108840, Russia
‡National Research University, Higher School of Economics, Moscow 101000, Russia
⊥Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the RAS, Moscow 117997, Russia
# Moscow Institute of Physics and Technology, Dolgoprudny, Moscow reg. 141700, Russia
§The University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555, USA
Abstract
In this work, we present a new approach to the detection of Troponin-T molecules in human blood serum. The approach includes: (1) a unique sample preparation method that allows selectively (using appropriate antibodies) to attach about 30 fluorescent molecules to each Troponin-T molecule, (2) optical microscopy of fluorescent molecules attached to the Troponin- T, which makes it possible to visualize in an optical microscope the diffuse movement of one Troponin-T molecule in human blood serum, (3) the use of carefully selected excitation laser wavelengths to minimize parasitic luminescence. As a result, we were able to demonstrate the detection of Troponin-T molecules 1000 times faster than the known approaches. The indicated detection limit was about 1 pg/ml - an order of magnitude better than existing methods.
Speaker
Pavel Melentiev
Institute of Spectroscopy
Russia
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