- Oblique Plane Microscopy (OPM) is an innovative fluorescence imaging technique that enables rapid, three-dimensional imaging of living specimens by illuminating and detecting the sample along a tilted (oblique) plane. It combines the advantages of light-sheet fluorescence microscopy with a single-objective design, allowing volumetric imaging without physically moving the sample or objective.
- The core principle of OPM is the use of an obliquely oriented light sheet that illuminates a tilted plane within the specimen. This light sheet is introduced through the primary objective at an angle, and the emitted fluorescence is collected by the same objective. A remote refocusing system, often involving secondary optics and cameras aligned to match the tilted illumination plane, reconstructs the optical sections into 3D images. This configuration allows for high-speed imaging of dynamic processes in living cells and tissues.
- One of the major benefits of OPM is its minimally invasive design. Since both excitation and detection occur through a single objective, it is compatible with standard microscope setups and sample holders, making it more accessible than traditional dual-objective light-sheet systems. Moreover, OPM achieves fast volumetric acquisition rates, ideal for capturing rapid biological events such as vesicle trafficking, calcium signaling, or cell migration.
- Additionally, OPM reduces photodamage by confining the excitation to a thin slice of the specimen, much like light-sheet microscopy, but without requiring perpendicular objectives or complex sample mounting. It is particularly useful for live-cell imaging, embryonic development studies, and high-throughput applications.
- In summary, Oblique Plane Microscopy offers a unique approach to fast and gentle 3D fluorescence imaging. By combining tilted light-sheet illumination with a single-objective detection system, it enables high-resolution, real-time imaging of biological processes in live specimens with minimal phototoxicity and mechanical disruption.
