A perspective on light sheet microscopy and imaging: Applications across the breadth of applied physics and biophysics

Abstract

Once in a while, a field of research comes across a technological breakthrough that drives it forward. This is something that seems apparent in optical fluorescence microscopy and imaging with the advent of light sheet technology. Over a span of a few years, this technology has revolutionized the field and has brought in the concept of sheet-based plane-selective imaging with near-diffraction-limited resolution. The technology has progressed further by its integration with existing imaging methods and super-resolution techniques. Unlike existing techniques that are predominantly point-illumination-based, light sheet technology offers selectivity, single-shot interrogation, a large field-of-view, and high-speed data acquisition. Qualitatively, this has resulted in an improved signal-to-background ratio, reduced scattering, improved contrast, and the ability to interrogate a large spectrum of specimens (from cells to mammals). Recent advances have shown the reach of this emerging technology in biological sciences (developmental biology and imaging flow cytometry) and applied physics (nanolithography, beam shaping, and optical traps). The growth and adaptability of this technique are well received by the research community. The technique is expected to grow at a steady rate and has the ability to overcome the issues faced by the existing point-based techniques. In this perspective, we look at the challenges of live imaging (for mammals and plants), the methods in applied physics, and the opportunities presented by recent advances.