In recent years, a revolution in the basic sciences has leveraged novel single molecule detection and manipulation tools to analyze biomolecules in unprecedented detail.
The University of Michigan's Single Molecule Analysis in Real-Time (SMART) Center aims to develop a university-wide initiative in this emerging field, encompassing techniques such as single molecule spectroscopy (SMS), single molecule imaging (SMI), laser tweezers (LT), and atomic force microscopy (AFM). This initiative seeks to unite basic scientists, engineers, and clinical researchers to address critical questions in medicine and nanotechnology, building on the momentum of the Michigan Lifescience Corridor (MLSC) Core Technology Alliance.
Single molecule methods offer detailed insights into molecular mechanisms and interactions, complementing bulk assays with information that traditional methods often miss. These approaches also underpin super-resolution imaging techniques like PALM/STORM.
The Value of Single Molecule Studies
By analyzing the behavior of individual particles, single molecule techniques reveal population characteristics obscured by ensemble averaging. They allow direct observation of:
- Molecular sub-populations
- Short-lived or transient states
- Rare molecular events
- Non-uniform kinetics
- Mechanical and thermodynamic details of mechanochemical events in individual molecules
Reviews of Supported Assays
- “Do-it-yourself guide: how to use the modern single-molecule toolkit” — Walter et al., Nature Methods (2008)
- “Biological mechanisms, one molecule at a time” — Tinoco and Gonzales, Genes and Development (2011)
- “A practical guide to single-molecule FRET” — Roy, Hohng, and Ha, Nature Methods (2008)
- “Single-molecule force spectroscopy: optical tweezers, magnetic tweezers and atomic force microscopy” — Neuman and Nagy, Nature Methods (2008)
Modalities and Measurands Overview
The SMART Center offers a range of single molecule study modalities, each with specific example measurands that illustrate the breadth of research capabilities:
- Single Molecule Fluorescent Imaging: Particle tracking, co-localization, stoichiometry quantification, and super-resolution imaging.
- Single Molecule FRET: Conformational changes, binding kinetics, and intra-/inter-molecular arrangements.
- Fluorescence Lifetime Imaging (FLIM): Background-resistant imaging and FRET analysis.
- Fluorescence Correlation Spectroscopy (FCS): Particle concentration, diffusion coefficients, and intermolecular binding kinetics.
- Force Spectroscopy: Protein force production, membrane tension, and intermolecular binding energy.
- Custom Assay Development and More: Tailored research solutions.
This structured breakdown emphasizes our commitment to advancing research in single molecule studies by providing state-of-the-art methodologies for diverse scientific inquiries.
