The ultimate goal of biological imaging is to be in a position to visualize systems that are increasingly physiologically relevant. A series of technological advances over the past two decades, such as the one in the near future, and the broad array of advanced research methods available means it is now possible to visualize many systems.

A crucial aspect of visualizing biological experiments is the effect of the imaging techniques that are used. It is necessary to achieve visualization that preserves the physiological integrity of the system being studied and reduces disturbances and the introduction of artifacts.

One of the most difficult aspects of biological imaging is reducing or, perhaps, eliminating phototoxicity and photodamage. Exposure to light, specifically the high intensities of light required for fluorescent imaging, could have a drastic effect on the function and well-being of organisms and living cells. 

Light Sheet Microscopy

Light sheet microscopy is a common term that describes an expanding class of planar illumination methods that have transformed the optical imaging of biological specimens. It was rendered feasible by isolating the illumination and detection of optical pathways to enable the use of the innovative technique of less damaging illumination.

The light sheet that is the foundation for the method is formed by laser light shaped into a hyperbolic "sheet" of illumination. As an alternative, an approximation to a light sheet can be achieved using a scanned beam. Detection is carried out along an axis that is different from that of the axis of illumination.

Applications of Light Sheet Microscopy

Light sheet microscopy is a virtually non-destructive and valuable imaging tool. It offers higher resolution and faster imaging speed compared to other non-destructive imaging methods, such as computerized tomography and magnetic resonance imaging. 

The high-resolution visualization of the sub-cellular structure is realized by using a plane of light to enable optical sectioning of a living organism or tissue.  Light sheet microscopy has advanced research in various fields by facilitating the rapid imaging of biological samples, including those larger than can be viewed with other microscopy methods, with higher resolution.

A light sheet microscopy is also a versatile tool for performing rapid, high-resolution quantitative mapping of the structure and function of large biological systems. It can be used to image the non-isomorphic variations in the body shape of the hydra as well as a clarified thick coronal slab of the human brain.

Imaging Of Intact Tissues

It has been demonstrated that the method allows rapid 2D and 3D imaging of intact tissues with the same level of detail as conventional pathology. For instance, it can enable quicker intra-operative evaluation of tumor-margin surfaces or volumetric assessment of optically cleared core-needle biopsies.

The Future of Light Sheet Microscopy

A light sheet microscopy is a valuable tool with the potential to perform not only rapid surface microscopy but also deep volumetric microscopy of specimens. Thanks to its non-destructive nature, it can be used to study a broad array of living biological systems. Moreover, it can allow live 3D imaging with the only minimal risk of photodamage.

Furthermore, the interpretation of the obtained data will be further enhanced by the development of visualization tools and pattern recognition software, thus enabling the generation of models for human physiology and disease.