New Microscope Harnesses Hybrid Technology for Advanced 3D Molecular Imaging

The adage “two heads are better than one” applies not only to teamwork but also to technological innovation in scientific research. This principle is exemplified in a groundbreaking hybrid microscope developed at the Marine Biological Laboratory (MBL), capable of simultaneously imaging the full 3D orientation and position of molecular ensembles within cells.

Revolutionizing Molecular Imaging

This advanced microscope, detailed in the current issue of Proceedings of the National Academy of Sciences, merges polarized fluorescence technology with dual-view light sheet microscopy (diSPIM). The combination allows researchers to observe and analyze proteins and other molecular structures in unprecedented detail.

Understanding Molecular Movements

Proteins within cells are highly dynamic, changing their 3D orientation in response to their environment. This movement is essential for their functions, including interaction with other molecules. The new microscope captures these 3D orientation changes, revealing aspects of cell biology that were previously difficult to study.

“Using this instrument, 3D protein orientation changes can be recorded. There’s real biology that might be hidden to you from just a position change of a molecule alone,”

— Talon Chandler, CZ Biohub San Francisco

Challenging Traditional Methods

Imaging the spindle of dividing cells, a significant challenge in microscopy, becomes feasible with this new technology. Traditional methods struggled with capturing accurate 3D information when the spindle plane was tilted. The hybrid microscope, however, can “correct” for these angles to provide clear images.

With traditional microscopy, including polarized light, you can study the spindle quite nicely if it’s in the plane perpendicular to the viewing direction. As soon as the plane is tilted, the readout becomes ambiguous.

— Rudolf Oldenbourg, Marine Biological Laboratory

The Development Journey

The idea for this innovative microscope emerged through a collaborative effort in 2016 at the MBL. Hari Shroff, then working at the National Institutes of Health (NIH) and now at HHMI Janelia, was using his custom-designed diSPIM microscope. This microscope, developed in collaboration with Abhishek Kumar, provides a dual view of samples, improving depth resolution and polarization control.

Overcoming Limitations

During discussions with Rudolf Oldenbourg, a senior scientist at MBL, they recognized the potential of combining diSPIM with polarized light microscopy. The diSPIM’s dual-view capability could solve challenges encountered with polarized light, particularly in efficiently illuminating samples.

“If we had two orthogonal views, we could sense polarized fluorescence along that direction much better,” Shroff explained. This realization led to the decision to integrate polarized fluorescence measurements into the diSPIM design.

Collaborative Innovation

Patrick La Rivière, a professor at the University of Chicago, whose lab specializes in computational imaging algorithms, joined the project. La Rivière brought his graduate student, Talon Chandler, to the MBL. Chandler’s doctoral thesis became focused on this hybrid microscope project, which he developed further in Oldenbourg’s lab.

The team included Shalin Mehta, also based at MBL, and they fitted the diSPIM with liquid crystals to manipulate and change the direction of input polarization.

“There was tons of cross-talk between the MBL, the University of Chicago, and the NIH, as we worked this through,” Chandler noted.

Future Applications and Improvements

The new microscope offers a glimpse into future possibilities in molecular imaging. Researchers aim to enhance the system’s speed to enable real-time observation of molecular changes in live samples. Additionally, the development of novel fluorescent probes could expand the variety of biological structures imaged with this technology.

Conclusion

This innovative hybrid microscope represents a significant leap forward in the world of molecular imaging. By combining polarized fluorescence and dual-view light sheet microscopy, scientists gain unprecedented insight into the complex, 3D world within our cells.

What Do You Think?

We’d love to hear your thoughts on this revolutionary advancement. Please leave your comments below, share your experiences, and join the conversation. Don’t forget to subscribe to our newsletter for the latest updates in scientific research and discoveries.

Spread the word and share this article on your social media platforms to engage more with the scientific community!