Light-sheet microscopy: a tutorial

This paper is intended to give a comprehensive review of light-sheet (LS) microscopy from an optics perspective. As such, emphasis is placed on the advantages that LS microscope configurations present, given the degree of freedom gained by uncoupling the excitation and detection arms. The new imaging properties are first highlighted in terms of optical parameters and how these have enabled several biomedical applications. Then, the basics are presented for understanding how a LS microscope works. This is followed by a presentation of a tutorial for LS microscope designs, each working at different resolutions and for different applications. Then, based on a numerical Fourier analysis and given the multiple possibilities for generating the LS in the microscope (using Gaussian, Bessel, and Airy beams in the linear and nonlinear regimes), a systematic comparison of their optical performance is presented. Finally, based on advances in optics and photonics, the novel optical implementations possible in a LS microscope are highlighted.Peer ReviewedPostprint (published version

Reconstruction of hidden 3D shapes using diffuse reflections

We analyze multi-bounce propagation of light in an unknown hidden volume and demonstrate that the reflected light contains sufficient information to recover the 3D structure of the hidden scene. We formulate the forward and inverse theory of secondary and tertiary scattering reflection using ideas from energy front propagation and tomography. We show that using careful choice of approximations, such as Fresnel approximation, greatly simplifies this problem and the inversion can be achieved via a backpropagation process. We provide a theoretical analysis of the invertibility, uniqueness and choices of space-time-angle dimensions using synthetic examples. We show that a 2D streak camera can be used to discover and reconstruct hidden geometry. Using a 1D high speed time of flight camera, we show that our method can be used recover 3D shapes of objects "around the corner"

Apparatus to control and visualize the impact of a high-energy laser pulse on a liquid target

We present an experimental apparatus to control and visualize the response of a liquid target to a laser-induced vaporization. We use a millimeter-sized drop as target and present two liquid-dye solutions that allow a variation of the absorption coefficient of the laser light in the drop by seven orders of magnitude. The excitation source is a Q-switched Nd:YAG laser at its frequency-doubled wavelength emitting nanosecond pulses with energy densities above the local vaporization threshold. The absorption of the laser energy leads to a large-scale liquid motion at timescales that are separated by several orders of magnitude, which we spatiotemporally resolve by a combination of ultra-high-speed and stroboscopic high-resolution imaging in two orthogonal views. Surprisingly, the large-scale liquid motion at upon laser impact is completely controlled by the spatial energy distribution obtained by a precise beam-shaping technique. The apparatus demonstrates the potential for accurate and quantitative studies of laser-matter interactions.Comment: Submitted to Review of Scientific Instrument

Diffractive oblique plane microscopy

Imaging of neuronal activity with fluorescent indicators is an important technique in neuroscience. However, it remains challenging to record volumetric image data at fast frame rates and good resolution. One promising technique to achieve this goal is light sheet microscopy (LSM), but the right angle configuration of the excitation and imaging system limits its application. Oblique plane microscopy (OPM), a variant of LSM, circumvents this limitation by exciting oblique planes and detecting the image through the same microscope objective lens. So far, these techniques have relied on the use of high numerical aperture (NA) detection objective lenses, which limits their field of view. Here we present an OPM technique that allows for the use of low NA objective lenses by redirecting the light with the help of a diffraction grating. The microscope maintains a micrometer-scale lateral resolution over a large addressable imaging volume of 3.3×3.0×1.0  mm3. We demonstrate its practicality by imaging the whole brain of larval and juvenile zebrafish