scholarly journals Tiling light sheet selective plane illumination microscopy using discontinuous light sheets

2019 ◽  
Vol 27 (23) ◽  
pp. 34472 ◽  
Author(s):  
Dongyue Wang ◽  
Yuxiao Jin ◽  
Ruili Feng ◽  
Yanlu Chen ◽  
Liang Gao
Keyword(s):  
Light Sheet ◽  
Selective Plane Illumination Microscopy

scholarly journals Tiling light sheet selective plane illumination microscopy using discontinuous light sheets

2018 ◽  
Author(s):  
Liang Gao
Keyword(s):  
Spatial Resolution ◽  
3D Imaging ◽  
High Spatial Resolution ◽  
Image Data ◽  
Image Plane ◽  
Light Sheet ◽  
Selective Plane Illumination Microscopy ◽  
Novel Method ◽  
Technical Details ◽  
Imaging Speed

AbstractTiling light sheet selective plane illumination microscopy (TLS-SPIM) improves 3D imaging ability of SPIM by using a real-time optimized tiling light sheet. However, the imaging speed decreases, and size of the raw image data increases proportionally to the number of tiling positions in TLS-SPIM. The decreased imaging speed and the increased raw data size could cause significant problems when TLS-SPIM is used to image large specimens at high spatial resolution. Here, we present a novel method to solve the problem. Discontinuous light sheets created by scanning coaxial beam arrays synchronized with camera exposures are used for 3D imaging to decrease the number of tiling positions required at each image plane without sacrificing the spatial resolution. We investigate the performance of the method via numerical simulation and discuss the technical details of the method.

scholarly journals A method to keep the excitation light sheet in focus in selective plane illumination microscopy

2017 ◽  
Author(s):  
Liang Gao
Keyword(s):  
3D Imaging ◽  
Light Sheet ◽  
Axial Position ◽  
Excitation Light ◽  
Selective Plane Illumination Microscopy

AbstractKeeping the excitation light sheet in focus is critical in selective plane illumination microscopy (SPIM) to ensure its 3D imaging ability. Unfortunately, an effective method that can be used in SPIM on general biological specimens to find the axial position of the excitation light sheet and keep it in focus is barely available. Here, we present a method to solve the problem. We investigate its mechanism and demonstrate its performance on a lattice light sheet microscope.

scholarly journals μSPIM: A Software Platform for Selective Plane Illumination Microscopy

2020 ◽  
Author(s):  
Daniel Saska ◽  
Paul Pichler ◽  
Chen Qian ◽  
Chrysia Pegasiou ◽  
Christopher L. Buckley ◽  
...  
Keyword(s):  
Optical Design ◽  
Light Sheet ◽  
Volumetric Imaging ◽  
Larval Zebrafish ◽  
Selective Plane Illumination Microscopy ◽  
Spatio Temporal ◽  
Set Up ◽  
Control And Synchronization ◽  
The Brain ◽  
Camera Shutter

AbstractSelective Plane Illumination Microscopy (SPIM) is a fluorescence imaging technique that allows volumetric imaging at high spatio-temporal resolution to monitor neural activity in live organisms such as larval zebrafish. A major challenge in the construction of a custom SPIM microscope is the control and synchronization of the various hardware components. Here we present a control toolset, μSPIM, built around the open-source MicroManager platform that has already been widely adopted for the control of microscopy hardware. Installation of μSPIM is relatively straightforward, involving a single C++ executable and a Java-based extension to Micro-Manager. Imaging protocols are defined through the μSPIM extension to Micro-Manager. The extension then synchronizes the camera shutter with the galvanometer mirrors to create a light-sheet that is scanned in the z-dimension, in synchrony with the imaging objective, to produce volumetric recordings. A key advantage of μSPIM is that a series of calibration procedures optimizes acquisition for a given set-up making it relatively independent of the optical design of the microscope, or the hardware used to build it. Two laser illumination arms can be used while also allowing for the introduction of illumination masks. μSPIM allows imaging of calcium activity throughout the brain of larval zebrafish at rates of 100 planes per second with single cell resolution as well as slower imaging to reconstruct cell populations, for example, in the cleared brains of mice.

scholarly journals Image quality guided smart rotation improves coverage in microscopy

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiaye He ◽  
Jan Huisken
Keyword(s):  
Image Quality ◽  
Light Sheet ◽  
Field Of View ◽  
Optical Performance ◽  
Sample Rotation ◽  
Optical Instruments ◽  
Selective Plane Illumination Microscopy ◽  
Data Volume ◽  
Optical Paths ◽  
Additional Hardware

AbstractFluorescence microscopy is an essential tool for biological discoveries. There is a constant demand for better spatial resolution across a larger field of view. Although strides have been made to improve the theoretical resolution and speed of the optical instruments, in mesoscopic samples, image quality is still largely limited by the optical properties of the sample. In Selective Plane Illumination Microscopy (SPIM), the achievable optical performance is hampered by optical degradations encountered in both the illumination and detection. Multi-view imaging, either through sample rotation or additional optical paths, is a popular strategy to improve sample coverage. In this work, we introduce a smart rotation workflow that utilizes on-the-fly image analysis to identify the optimal light sheet imaging orientations. The smart rotation workflow outperforms the conventional approach without additional hardware and achieves a better sample coverage using the same number of angles or less and thereby reduces data volume and phototoxicity.

scholarly journals Selective plane illumination microscopy on a chip

Lab on a Chip ◽  
2016 ◽  
Vol 16 (9) ◽  
pp. 1556-1560 ◽  
Author(s):  
Petra Paiè ◽  
Francesca Bragheri ◽  
Andrea Bassi ◽  
Roberto Osellame
Keyword(s):  
High Throughput ◽  
Light Sheet ◽  
3D Reconstructions ◽  
Selective Plane Illumination Microscopy ◽  
Large Populations ◽  
On Chip

A high-throughput on-chip light sheet microscope allowing 3D reconstructions of large populations of samples, even with standard microscopes, is presented.

scholarly journals Multidirectional digital scanned light-sheet microscopy enables uniform fluorescence excitation and contrast-enhanced imaging

2018 ◽  
Author(s):  
Adam K. Glaser ◽  
Ye Chen ◽  
Chengbo Yin ◽  
Linpeng Wei ◽  
Lindsey A. Barner ◽  
...  
Keyword(s):  
Scattered Light ◽  
Light Sheet ◽  
Line Detection ◽  
Fluorescence Excitation ◽  
Light Sheet Microscopy ◽  
Contrast Enhanced ◽  
Selective Plane Illumination Microscopy ◽  
Light Sheet Fluorescence Microscopy ◽  
Enhanced Imaging ◽  
Imaging Plane

AbstractLight-sheet fluorescence microscopy (LSFM) has emerged as a powerful method for rapid and optically efficient 3D microscopy. Initial LSFM designs utilized a static sheet of light, termed selective plane illumination microscopy (SPIM), which exhibited shadowing artifacts and deteriorated contrast due to light scattering. These issues have been addressed, in part, by multidirectional selective plane illumination microscopy (mSPIM), in which rotation of the light sheet is used to mitigate shadowing artifacts, and digital scanned light-sheet microscopy (DSLM), in which confocal line detection is used to reject scattered light. Here we present a simple passive multidirectional digital scanned light-sheet microscopy (mDSLM) architecture that combines the benefits of mSPIM and DSLM. By utilizing an elliptical Gaussian beam with increased angular diversity in the imaging plane, mDSLM provides shadow-free contrast-enhanced imaging of fluorescently labeled samples.One Sentence SummaryGlaser et al. describe a light-sheet microscopy architecture that enables passive multidirectional illumination with confocal line detection to enable both uniform fluorescence excitation and contrast-enhanced imaging of fluorescently labeled samples.

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