Upgraded cellular and volumetric imaging available to users at UCLA facility

Spinning disk confocal microscope brings speed, motion and color to biological discovery, now online at CNSI Tech Center

Melanoma cells labeled growing to confluency over 48 hours. Cells can be seen detaching from the surface, dividing, then reattaching. (Image courtesy: ALMS/CNSI)

To expand the understanding of fundamental processes in living things, researchers often need to capture images of biological dynamics spooling out over fractions of a second or volumetric data over periods of days.

Investigators looking to create knowledge about such difficult-to-capture phenomena have a newly added resource in an upgraded spinning disk confocal microscope, now available at the Advanced Light Microscopy and Spectroscopy (ALMS) laboratory. A Technology Center of the California NanoSystems Institute at UCLA, ALMS provides a unique collection of high-end instruments for fluorescence-based imaging and measurements, as well as custom devices for gathering image data from small animal models.

A longtime standard now upgraded with state-of-the-art lasers, the Yokogawa Confocal Scanner Unit offers high-speed acquisition of 3D fluorescence data — up to 200 frames per second. The ability to delicately produce time-lapse recordings makes it possible to image the motion of live cells and conduct longitudinal studies over the course of days.

Two spinning disks in the microscope respectively have about 20,000 microlenses and a matching number of pinholes to focus light and provide optical sectioning with high photonic efficiency. By rotating them at up to 5,000 rpm, the device rapidly scans the field of view through these disks to sensitively detect fluorescence signal plane-by-plane, while minimizing bleaching and toxicity to maintain sample health.

Jing Wang, a PhD candidate in bioengineering, has used the instrument for macrostructure analysis.

“The spinning disk provides fast, high-resolution images of the living zebrafish heart, allowing us to study the flow-structure interaction during cardiac development,” he said.

Beating heart of a zebrafish embryo. Volume reconstructed with a phase-matching algorithm from images taken on the spinning disk. (Image courtesy: Jing Wang, Bioengineering; ALMS/CNSI)

There is a wide range of magnification powers, up to 100× for subcellular details. The filter wheel and acousto-optic tunable filters synchronize for fast color-switching, and users can image up to four dyes in 3D with excitation laser lines of wavelengths at 405, 488, 561 and 640 nanometers, as well as matching emission filters.

Combined with other ALMS equipment such as the Tokai Hit stage-top incubator, which provides a controlled environment for live growth, the spinning disk confocal microscope presents the capacity to examine live or fixed human tissue and animal cells.

The microscope also enables molecular studies conducted by Dylan Valencia, a PhD candidate in chemistry and biochemistry.

“The high frame rates on the spinning disk confocal allow me to gain information on calcium cation transients in neonatal rat cardiomyocytes,” he said. “The customizability of the UI provides the flexibility to take images at different time intervals. And the incubation system allows great control over the temperature of my live cells.”

Another key feature is the microscope’s ease of use. With various stage inserts to accommodate common sample mounting and a user-friendly interface, it has been described as a turnkey system that allows scientists and engineers to quickly proceed to the important task of acquiring vital data.

The spinning disk confocal microscope is available to all users, including interested industry and business partners, academic investigators from other campuses, and UCLA researchers, faculty and students. For general information about the spinning disk confocal microscope, prospective users may contact alms@cnsi.ucla.edu. Those interested in project consultation and proof-of-concept should reach out to Laurent Bentolila, director of ALMS, at lbento@chem.ucla.edu. Training and assistance can be arranged with Brian Jeong, research and development engineer, at bjeong@cnsi.ucla.edu.