This method of super-resolution microscopy uses subtracted interleaved solid and doughnut-shaped laser pulses to improve spatial resolution, resulting in real-time imaging of live microscopic organisms. The live cell imaging market has an estimated worth of $3.57 billion and is expected to grow due to increased government funding and the demand for high-content screening microscopy techniques in drug discovery. Conventional super-resolution microscopy requires high light dosage or special dyes to achieve improved spatial resolution, which can lead to damage in live cells during the imaging process. Clemson University researchers have developed a method of super-resolution microscopy using low power laser pulses that is compatible with an unrestricted menu of dyes or chromophores, allowing for imaging of live samples without causing damage to the cellular architecture. This innovative use of low power settings makes this technology promising for the future of biological research.
Application Stage of Development
Real-time, live cell imaging Proof of concept
Advantages
• Pulse-to-pulse deduction with multiplexed two modes of pulse illumination, improving spatial resolution
at least twice over the diffraction limit
• Single wavelength laser is used, simplifying the equipment traditionally needed for super-resolution microscopy.
• Lower excitation power compared to Stimulated Emission Depletion microscopy, preventing cell damage
Technical Summary
In this method of super-resolution microscopy, interleaved solid and doughnut-shaped laser pulses are used to excite the biological sample, and the signal generated from each pulse is recorded. The signals between neighboring pulses are subtracted to effectively yield a shrunken point spread function, providing improved spatial resolution. The improvement factor is over two without deconvolution. This method uses only one laser at a single wavelength, which removes the need for high powered lasers used by other super-resolution microscopy techniques. The reduced power allows for imaging of live samples without damaging them. Low power pulsed lasers on the market cover a wide range of wavelengths, spanning from UV to near infrared. This allows for an unlimited selection of dyes or chromophores to use during the imaging process.
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Inventor: Dr. Tong Ye
Patent Type: Provisional
Serial Number: 62/482,251
CURF Ref No: 2017-048