Visualizing Whole-Brain Activity and Development at the Single-Cell Level Using Light-Sheet Microscopy

Keller, Philipp J.; Ahrens, Misha B.

doi:10.1016/j.neuron.2014.12.039

Cited by 224 publications

(187 citation statements)

References 101 publications

(169 reference statements)

Supporting

Mentioning

185

Contrasting

Unclassified

Order By: Relevance

“…Nowadays, complementary metal oxide semiconductor (cMOS) cameras are used to ensure a detection of the entire field at high speed and high resolution, way beyond the detection obtained with scanning technologies. Thus, owing to a reduced acquisition time as well as the simultaneous and selective illumination of the detection plane, LSFM significantly decreases the phototoxicity and photobleaching compared to classic confocal microscopy (Keller and Ahrens, 2015;Lim et al, 2014;Pampaloni et al, 2015;Weber and Huisken, 2011); it is therefore highly useful in imaging dynamic events that take place in small and transparent organisms and embryos. In this section, we briefly focus on the two main technological strategies that have been developed to achieve this specific illumination, single plane illumination microscopy (SPIM) and digitally scanned light-sheet microscopy (DSLM).…”

Section: Light-sheet Fluorescence Microscopy (Lsfm)mentioning

confidence: 99%

Seeing is believing: multi-scale spatio-temporal imaging towards in vivo cell biology

Follain

Mercier

Osmani

et al. 2016

Journal of Cell Science

View full text Add to dashboard Cite

Life is driven by a set of biological events that are naturally dynamic and tightly orchestrated from the single molecule to entire organisms. Although biochemistry and molecular biology have been essential in deciphering signaling at a cellular and organismal level, biological imaging has been instrumental for unraveling life processes across multiple scales. Imaging methods have considerably improved over the past decades and now allow to grasp the inner workings of proteins, organelles, cells, organs and whole organisms. Not only do they allow us to visualize these events in their most-relevant context but also to accurately quantify underlying biomechanical features and, so, provide essential information for their understanding. In this Commentary, we review a palette of imaging (and biophysical) methods that are available to the scientific community for elucidating a wide array of biological events. We cover the most-recent developments in intravital imaging, light-sheet microscopy, superresolution imaging, and correlative light and electron microscopy. In addition, we illustrate how these technologies have led to important insights in cell biology, from the molecular to the whole-organism resolution. Altogether, this review offers a snapshot of the current and state-of-the-art imaging methods that will contribute to the understanding of life and disease.

show abstract

Section: Light-sheet Fluorescence Microscopy (Lsfm)mentioning

confidence: 99%

Seeing is believing: multi-scale spatio-temporal imaging towards in vivo cell biology

Follain

Mercier

Osmani

et al. 2016

Journal of Cell Science

View full text Add to dashboard Cite

show abstract

“…[1][2][3] LSFM is fundamentally characterized by setting a separated plane illumination orthogonal to the wide-¯eld detection, thus enabling optical sectioning inside the samples with sharp contrast and high axial resolution superior to conventional microscopes. [4][5][6][7] In the past decade, besides the basic Gaussian light-sheet generated with a cylindrical lens, LSFM have derived a series of structural variants, including multidirectional SPIM (mSPIM), Digital Scanned Laser Light-sheet Fluorescence Microscopy (DSLM), Bessel beam plane illumination microscopy and so on.…”

Section: Introductionmentioning

confidence: 99%

Three-dimensional, isotropic imaging of mouse brain using multi-view deconvolution light sheet microscopy

Liu

Nie

et al. 2017

J. Innov. Opt. Health Sci.

View full text Add to dashboard Cite

We present a three-dimensional (3D) isotropic imaging of mouse brain using light-sheet°uo-rescent microscopy (LSFM) in conjunction with a multi-view imaging computation. Unlike common single view LSFM is used for mouse brain imaging, the brain tissue is 3D imaged under eight views in our study, by a home-built selective plane illumination microscopy (SPIM). An output image containing complete structural information as well as signi¯cantly improved resolution ($4 times) are then computed based on these eight views of data, using a bead-guided multi-view registration and deconvolution. With superior imaging quality, the astrocyte and pyramidal neurons together with their subcellular nerve¯bers can be clearly visualized and segmented. With further including other computational methods, this study can be potentially scaled up to map the connectome of whole mouse brain with a simple light-sheet microscope.

show abstract

“…LSFM enables imaging at high time resolution, while inducing less phototoxicity than laser scanning or spinning disc confocal microscopes 6,7 . Nowadays, there are already many implementations of the basic light sheet illumination principle and it has been used to image a large variety of specimens and processes previously inaccessible to researchers [8][9][10][11] .…”

Section: Introductionmentioning

confidence: 99%

Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development

Icha

Schmied

Sidhaye

et al. 2016

JoVE

View full text Add to dashboard Cite

Light sheet fluorescence microscopy (LSFM) is gaining more and more popularity as a method to image embryonic development. The main advantages of LSFM compared to confocal systems are its low phototoxicity, gentle mounting strategies, fast acquisition with high signal to noise ratio and the possibility of imaging samples from various angles (views) for long periods of time. Imaging from multiple views unleashes the full potential of LSFM, but at the same time it can create terabyte-sized datasets. Processing such datasets is the biggest challenge of using LSFM. In this protocol we outline some solutions to this problem. Until recently, LSFM was mostly performed in laboratories that had the expertise to build and operate their own light sheet microscopes. However, in the last three years several commercial implementations of LSFM became available, which are multipurpose and easy to use for any developmental biologist. This article is primarily directed to those researchers, who are not LSFM technology developers, but want to employ LSFM as a tool to answer specific developmental biology questions.Here, we use imaging of zebrafish eye development as an example to introduce the reader to LSFM technology and we demonstrate applications of LSFM across multiple spatial and temporal scales. This article describes a complete experimental protocol starting with the mounting of zebrafish embryos for LSFM. We then outline the options for imaging using the commercially available light sheet microscope. Importantly, we also explain a pipeline for subsequent registration and fusion of multiview datasets using an open source solution implemented as a Fiji plugin. While this protocol focuses on imaging the developing zebrafish eye and processing data from a particular imaging setup, most of the insights and troubleshooting suggestions presented here are of general use and the protocol can be adapted to a variety of light sheet microscopy experiments.

show abstract

Visualizing Whole-Brain Activity and Development at the Single-Cell Level Using Light-Sheet Microscopy

Cited by 224 publications

References 101 publications

Seeing is believing: multi-scale spatio-temporal imaging towards in vivo cell biology

Seeing is believing: multi-scale spatio-temporal imaging towards in vivo cell biology

Three-dimensional, isotropic imaging of mouse brain using multi-view deconvolution light sheet microscopy

Using Light Sheet Fluorescence Microscopy to Image Zebrafish Eye Development

Contact Info

Product

Resources

About