Abstract
Researchers at the University of Central Florida have
developed a line-scanning confocal fluorescence technology that scientists can
use to capture high-resolution, 3D, live-cell images while minimizing the
effects of photobleaching and photodamage to target molecules. By providing a higher
signal-to-noise-ratio (SNR) and lower excitation light intensity, the UCF multiple
line-scanning confocal microscopy allows
for longer observation time of fluorescent structures and reduces the effects
of photobleaching and photodamage. Overall, the design produces faster,
high-resolution images more efficiently than traditional line-scanning
microscopy while maintaining deep optical sectioning capability and
single-molecule sensitivity.
Technical Details
The UCF technology consists of a dual-inclined beam
line-scanning (2iLS) confocal microscope apparatus and method. The microscope
uses parallel excitation beams, each having a focused line shape that scans
over a fluorescent sample. The sample emits fluorescence that is spatially
filtered and detected by an array detector. In addition to two beams, the
technology design can accommodate multiple beams (such as four, six or eight). Many applications, including tissue imaging and high-throughput imaging, can easily incorporate the technology.
As shown in the figure, the microscope design can employ parallelized dual beams with inclined
illumination to lower the excitation intensity. The detector/camera assembly is
a scientific complementary metal-oxide
semiconductor (sCMOS) camera
supporting a rolling shutter mode. The assembly includes two electrical slits
to ensure the straightforward
implementation of simultaneous confocal detection. Experimental results showed
that the imaging method enables a two-fold longer observation time in
single-molecule imaging and immunofluorescence imaging compared to traditional
line-scanning microscopy.
Partnering Opportunity
The research team is looking for partners to develop the
technology further for commercialization.
Stage of Development
Prototype available.
Benefit
Provides higher signal-to-noise-ratio and less photobleaching/photodamageMarket Application
Single-molecule fluorescence imaging, high-throughput imaging, super-resolution imaging, tissue imagingHigh-resolution fluorescence microscope for biomedical applicationsPublications
Low-photobleaching line-scanning confocal microscopy using dual-inclined beams, J Biophotonics, 2019 May 20:e201900075. DOI: 10.1002/jbio.201900075
Brochure
