- Swept Confocally-Aligned Planar Excitation (SCAPE) microscopy is an advanced light-sheet imaging technique designed to rapidly capture high-resolution, volumetric data from live, dynamic biological samples without the need for sample translation or mechanical scanning. SCAPE combines the speed and low phototoxicity of light-sheet microscopy with a unique optical geometry that enables real-time, three-dimensional imaging using a single objective lens.
- The key innovation in SCAPE microscopy lies in its use of a scanned oblique light sheet, which is swept across the sample while fluorescence is collected through the same objective at a tilted angle. The scanning mirror both sweeps the excitation light and simultaneously de-scans the emitted fluorescence to keep it optically aligned with a stationary camera detector. This synchronization allows for continuous, high-speed imaging of a 3D volume, often exceeding tens of volumes per second, depending on the field of view and resolution.
- SCAPE eliminates the need to physically move the specimen or objective for volumetric imaging, making it particularly well-suited for live, freely moving organisms or rapidly changing processes such as neural activity, blood flow, embryonic development, and cell migration. Its single-objective configuration also simplifies sample mounting and improves compatibility with traditional biological setups.
- Additionally, SCAPE microscopy offers the following advantages:
- Minimized phototoxicity: Only a thin plane is illuminated at any given time, reducing energy exposure to the rest of the sample.
- High temporal resolution: Enables real-time 4D imaging (3D over time), crucial for capturing transient biological phenomena.
- Flexible sample compatibility: Suitable for specimens ranging from single cells to entire small organisms such as zebrafish, Drosophila larvae, and C. elegans.
- In summary, SCAPE microscopy is a transformative imaging modality that provides fast, volumetric, and minimally invasive imaging of living systems. Its unique approach to scanning and detection allows researchers to study complex, dynamic processes in their native 3D environments with unprecedented temporal and spatial resolution.
