Light-sheet microscopy with length-adaptive Bessel beams

Abstract

In light-sheet microscopy, a confined layer in the focal plane of the detection objective is illuminated from the side. The illumination light-sheet usually has a constant beam length independent of the shape of the biological object. Since the thickness and the length of the illumination light-sheet are coupled, a tradeoff between resolution, contrast and field of view has to be accepted. Here we show that scanned Bessel beams enable object adapted tailoring of the light-sheet defined by its beam length and position. The individual beam parameters are obtained from automatic object shape estimation by low-power laser light scattered at the object. Using Arabidopsis root tips, cell clusters and zebrafish tails, we demonstrate that Bessel beam light-sheet tailoring leads to a 50% increase in image contrast and a 50% reduction in photobleaching. Light-sheet tailoring requires only binary amplitude modulation, therefore allowing a real time illumination adaptation with little technical effort in the future.

Cell and tissue manipulation with ultrashort infrared laser pulses in light-sheet microscopy

“…In line with recent innovations to shape the light-sheet 47,48 and correct image aberrations 49 , an interesting future development of our integrated framework is to combine these innovations with laser manipulation to better resolve and target structures in the 3D space or in time.…”

Wide field light-sheet microscopy with lens-axicon controlled two-photon Bessel beam illumination

“…Applications of the tunable DSLM demonstrated imaging analysis of medaka development from embryos to juveniles in a single scheme by adapting the beam extent to the samples. A previous study 39 reported length-adaptive Bessel beams in which the focal position and beam length of Bessel beams are automatically adapted to the samples by modulating SLM. This approach can cover samples lying over the entire FOV, but only applies to thin samples that do not fill the whole FOV.…”

Metasurfaces for biomedical applications: imaging and sensing from a nanophotonics perspective

“…Metasurface-based structured beams such as vortex beams [175,176], Bessel beams (see Figure 10c) [177] and Bessel beams on a fiber tip [178]. This is important for STED microscopy [179], light-sheet microscopy [180], and two-photon microscopy [181]. Fourthly, conformal metasurfaces [64,182] are another intriguing area of exploration.…”