Single-molecule detection provides researchers with a unique method to probe kinetics of biomolecules in their native environment, without the need to synchronize the molecular states. However, current single-molecule measurements of DNA hybridization kinetics are mostly performed on a surface or inside an electrokinetic trap, which are not physiologically relevant conditions. Recently we demonstrate a time-resolved, 3D single-molecule tracking (3D-SMT) method that that can follow individual DNA molecules diffusing inside a mammalian cell and observe multiple annealing and melting events on the same molecules. By comparing the hybridization kinetics of the same DNA strand in vitro, we found the association constants can be 13- to 163-fold higher in the molecular crowding cellular environment. In contrast to other confocal-feedback 3D single-particle tracking demonstrations, we tracked single DNA reporter strands inside a live cell and measured their annealing-melting kinetics. Although camera-based techniques combined with point-spread function engineering can achieve 3D tracking in live cells, they do not offer any lifetime monitoring capability that can be used to reveal the molecular binding kinetics...
Recent Publications
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Y.-I. Chen, Y.-J. Chang, S.-C. Liao, T.D. Nguyen, J. Yang, Y.-A. Kuo, S. Hong, H.-C. Li, Y.-L. Liu, H.G. Rylander III, S.R. Santacruz, T.E. Yankeelov, and H.-C. Yeh, "Generative adversarial network enables rapid and robust fluorescence lifetime image analysis in live cells", Communications Biology 5,18, 2022.
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J. Yang, T. Davis, A.S. Kazerouni, Y.-I. Chen, M.J. Bloom, H.-C. Yeh, T.E. Yankeelov and J. Virostko, "Longitudinal FRET imaging of glucose and lactate dynamics and response to therapy in breast cancer cells", Molecular Imaging and Biology, 2021 .
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Y.-A. Kuo, C. Jung, Y.-A. Chen, H.-C. Kuo, O.S. Zhao, T.D. Nguyen, J.R. Rybarski, S. Hong, Y.-I. Chen, D.C. Wylie, J.A. Hawkins, J.T. Petty, I.J. Finkelstein and H.-C. Yeh, "Massively parallel selection of NanoCluster Beacons", bioRxiv, 2021.
News
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May 9, 2022
Nina recieved 2022 Outstanding Dissertation Award from UT. Congratulations!
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March 23, 2022
Michael gave an oral presentation on "Revealing nanocluster beacon design rules using an NGS chip" at the ACS Spring conference.
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March 22, 2022
Manasa Sripati won $1,000 Undergraduate Research Fellowship. Congratulations!
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February 27, 2022
Manasa Sripati and her teammates won the 2nd place in South-Southwest Medical Device Make-a-thon, hosted by the University of Texas at San Antonio, and awarded $300 prize. Congratulations!
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February 21, 2022
Ted gave a poster presentation on "Full-spectrum multiphoton autofluorescence imaging with temporal focusing" at the BPS conference.
Contact
Tim Yeh, Ph.D. (Hsin-Chih Yeh 葉信志)
Associate Professor
Department of Biomedical Engineering
University of Texas at Austin
107 W. Dean Keeton Street Stop C0800
Austin, TX 78712-1801
Office: BME 5.202C
Phone: (512) 471-7931
Email: tim.yeh@austin.utexas.edu