2018-718 Transverse Sheet Illumination Microscopy (TranSIM)

SUMMARY

UCLA researchers in the Department of Physics and Astronomy and at the California NanoSystems Institute have developed a highly customizable single objective microscopy technique which exhibits micron resolution in three dimensions, near millisecond temporal resolution (5 ms) and gentle illumination, allowing for the simultaneous imaging of multiple planes within a sample.

BACKGROUND

Confocal fluorescence microscopy is an optical imaging technique typically associated with high spatial resolution (~0.5 μm), but relatively slow temporal resolution (~100 ms) for volumetric scales. Currently, confocal microscopes operate by scanning a single plane, usually with slow (10 volumes/sec) scan rates, followed by sensing and deconvolution. The number of planes these microscopes can scan and remap is limited by the physical size of the sensing units, inherently restricting the technique. Therefore, a high-resolution microscopy method that can tackle imaging of volumetric samples at higher scan rates and across multiple planes would greatly augment the utility of this technology and accelerate imaging throughput.

INNOVATION

UCLA researchers in the Department of Physics and Astronomy and at the California NanoSystems Institute have developed a method for transverse sheet illumination microscopy that simultaneously allows for imaging of multiple z-planes while enhancing the temporal resolution of a single plane. The gentle illumination strategy prevents deleterious sample degradation, an important factor when imaging biological samples. Importantly, the intended scan rate of 200 volumes/sec significantly enhances the speed of microscopy compared to currently available spinning disk confocal microscopes and other lightsheet geometries. Furthermore, the system is scalable as evidenced by the cyclic nature of the plane separation technique. Overall, the TranSIM system is optimized for both spatial and temporal resolution, allowing for unprecedented dynamic investigations of ultra-fast biological processes such as neuronal transmission, action potential, brain activity and biochemical pathways.

POTENTIAL APPLICATIONS

• Biological samples
• Cell imaging
• Imaging of biochemical pathways
• Neuroscience
• Behavioral sciences
• Brain activity

ADVANTAGES

• Deconstruction of planes
• Construction of multiple planes into one
• Near millisecond temporal resolution (5 ms)
• Micron resolution in large three dimensions (460 μm x 682 μm x 160 μm)

DEVELOPMENT TO DATE

First successful demonstration of prototype