Highly Inclined Swept Tile (HIST) Microscope for Advanced Real-Time, Single-Molecule Imaging

Technology #34127

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Figure 1. An example setup of the UCF Highly Inclined Swept Tile (HIST) microscope system. CL1-CL2 are cylindrical lenses; DM is a dichroic mirror; GM is a galvo mirror; IP is the imaging plane; L1-L4 are relay lenses; M is a mirror; SMF is a single-mode fiber; TL is a tube lens.
Categories
Researchers
Kyu Young Han, Ph.D.
External Link (www.creol.ucf.edu)
Jialei Tang
Managed By
John Miner
Assistant Director 407.882.1136
Patent Protection

US Patent Pending US20190331904A1
Publications
Extended field-of-view single-molecule imaging by highly inclined swept illumination
Optica , Vol. 5, No. 9 September 2018
A Guide to Build a Highly Inclined Swept Tile Microscope for Extended Field-of-view Single-molecule Imaging
J. Vis. Exp. , (146), e59360, doi:10.3791/59360 (2019)

New fluorescence microscopy system enables super-resolution imaging of single molecules across a 40x wider field of view.

Researchers at the University of Central Florida have invented a fluorescence microscope that provides better real-time imaging of single molecules than conventional fluorescence technologies such as HILO (highly inclined and laminated optical sheet) microscopy. By enabling a much higher signal-to-background-ratio and a wider field-of-view (FOV), the UCF Highly Inclined Swept Tile (HIST) microscope delivers thinner illumination with greater than 40x FOV; thus providing clearer visualization of single molecules across a >130µm x 130µm FOV. With such capabilities, the innovation can benefit applications such as super-resolution imaging, single-molecule tracking, and smFISH-based high-throughput gene expression profiling.

Technical Details

The UCF HIST technology is a fluorescence microscope that uses a highly inclined tile beam to scan over a biological sample object.  The system spatially filters fluorescence emission from the sample through a programmable confocal slit into an sCMOS camera supporting a rolling shutter mode. The tile beam is synchronously swept with the readout of the camera to facilitate the rejection of background. The system provides for decoupling of the total imaging area from the beam thickness, which solely depends on the width of the tile beam, enabling a thinner illumination (high sectioning capability) and larger FOV for video-rate, live-cell imaging. The technology can be easily implemented onto a standard inverted microscope.

Partnering Opportunity

The research team is looking for partners to develop the technology further for commercialization.

Stage of Development

Prototype available.

Benefits

  • Higher signal-to-background-ratio
  • Wider FOV compared to current techniques
  • Provides high contrast images, high resolution, fast imaging speed, single-molecule sensitivity

Applications

  • High-resolution fluorescence microscopy for biomedical applications
  • Single-molecule fluorescence imaging
  • High-throughput transcriptomics
  • Super-resolution fluorescence imaging

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34128 Multiple Inclined Beam Line-Scanning Imaging Apparatus, Methods and Applications