The light-sheet microscopy revolution

Abstract

This paper reviews the rapid advances that have been made in one form of optical biological imaging in the last decade, namely that of light sheet microscopy. Although the concept was originally presented over one hundred years ago, at the time it was a methodology that lacked the technology to really make it a viable tool for practical everyday imaging in the biologist’s laboratory. However, since its re-discovery, it has started to transform in vivo and increasingly intact organ imaging in a number of areas of biology. This review looks back at the beginning of the method and then the crucial role that modern optical technology, frequently developed for other fields, has played in advancing the instrumentation. This paper will also look at the OpenSPIM route that was developed whereby, through the purchase of a few optical components, researchers have been able to develop their own bespoke instruments and we consider if this may be a route forward for the rapid development of other technological breakthroughs.

Multi-photon attenuation-compensated light-sheet fluorescence microscopy

“…x axis (µm) x axis (µm) x axis (µm) (a) (b) (c) (2) (2) (2) quired, and increases in SNR between 15 − 45% and 5 − 25% in the NDF-and SLM-based attenuation-compensation cases were observed at a depth of 200 µm.…”

“…Light-sheet fluorescence microscopy (LSFM) has transformed the field of imaging in recent years, owing to its optical sectioning capabilities, resulting in fast, highly resolved images with significantly reduced photo-bleaching and photo-toxicity [1,2]. Propagation-invariant light fields, such as Airy and Bessel beams, have been employed in LSFM not only because of their pseudo-nondiffracting properties which enables them to retain their transverse profile on propagation, but also due to their self-healing capabilities on interaction with obstacles during propagation [3][4][5][6].…”

Combined Atomic Force Microscope and Volumetric Light Sheet System for Mechanobiology

“…These goals necessitate integrating high spatial and temporal resolution volumetric imaging methods with high resolution force acquisition. Light sheet florescence microscopy (LSFM) enables acquisition of volumetric, multicolor time series at a high resolution and frame rate with low background fluorescence and low phototoxicity [4][5][6] . Among single cell force methods 7 , Atomic Force Microscopy (AFM) 8 is unique in combining a large force range (10 -11 -10 -6 N) that enables molecular-scale to tissue-level mechanics measurements, with high bandwidth temporal resolution (s) and sub-nanometer spatial control.…”

Mapping the microbiome location helps elucidate its role

“…Although FISH-based methods have provided unprecedented images of the microbiome, they only offer a snapshot in time. Raghuveer Parthasarathy, a biophysicist at the University of Oregon, instead uses a technique called light-sheet microscopy (7), which allows him to watch how a microbiome changes in space and time within a living gut.…”