Volumetric Imaging Gets Faster With New Microscopy Tech

Increasing the acquisition speed of three-dimensional volumetric imaging is essential – particularly in biological imaging – to unveil specimens’ structural dynamics and functionalities in detail. In conventional laser scanning fluorescence microscopy, volumetric images are constructed from optical sectioning images sequentially acquired by changing the observation plane, limiting the acquisition speed.

Researchers have developed a novel method to realize volumetric imaging from two-dimensional raster scanning of a light needle spot without sectioning, even in the traditional framework of laser scanning microscopy. The researchers simultaneously captured information from multiple axial planes using wavefront engineering for fluorescence signals. The researchers surveyed the entire depth range while maintaining spatial resolution.

This technique is applied to real-time and video-rate three-dimensional tracking of micrometer-sized particles and the prompt visualization of thick fixed biological specimens, offering substantially faster three-dimensional imaging.

The visualization of the three-dimensional (3D) structures of biological specimens is vital to examine their detailed behavior and functionality in vivo. Laser scanning microscopy has been extensively employed for this purpose owing to its optical sectioning ability accomplished by confocal detection using a small pinhole or multi-photon excitation processes for fluorescence specimens. However, 3D image acquisition using these point-scanning-based methods is, in principle, based on the image stacking of multiple images by changing the observation plane, which takes some time as the observation depth increases.


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