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Single-molecule and single-particle tracking experiments are typically unable to resolve fine details of thermal motion at short timescales where trajectories are continuous. We show that, when a diffusive trajectory [Formula: see text] is sampled at finite time intervals δt, the resulting error in measuring the first passage time to a given domain can exceed the time resolution of the measurement by more than an order of magnitude. Such surprisingly large errors originate from the fact that the trajectory may enter and exit the domain while being unobserved, thereby lengthening the apparent first passage time by an amount that is larger than δt. Such systematic errors are particularly important in single-molecule studies of barrier crossing dynamics. We show that the correct first passage times, as well as other properties of the trajectories such as splitting probabilities, can be recovered via a stochastic algorithm that reintroduces unobserved first passage events probabilistically.
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Top-down machine learning approach for high-throughput single-molecule analysis
Single-molecule approaches provide enormous insight into the dynamics of biomolecules, but adequately sampling distributions of states and events often requires extensive sampling. Although emerging experimental techniques can generate such large datasets, existing analysis tools are not suitable to process the large volume of data obtained in high-throughput paradigms. Here, we present a new analysis platform (DISC) that accelerates unsupervised analysis of single-molecule trajectories. By merging model-free statistical learning with the Viterbi algorithm, DISC idealizes single-molecule trajectories up to three orders of magnitude faster with improved accuracy compared to other commonly used algorithms. Further, we demonstrate the utility of DISC algorithm to probe cooperativity between multiple binding events in the cyclic nucleotide binding domains of HCN pacemaker channel. Given the flexible and efficient nature of DISC, we anticipate it will be a powerful tool for unsupervised processing of high-throughput data across a range of single-molecule experiments.
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- Award ID(s):
- 1856518
- PAR ID:
- 10545470
- Publisher / Repository:
- eLife
- Date Published:
- Journal Name:
- eLife
- Volume:
- 9
- ISSN:
- 2050-084X
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.7554/eLife.53357
