Adaptive light-sheet microscopy for long-term, high-resolution imaging in living organisms

2016 ◽  
Vol 34 (12) ◽  
pp. 1267-1278 ◽  
Author(s):  
Loïc A Royer ◽  
William C Lemon ◽  
Raghav K Chhetri ◽  
Yinan Wan ◽  
Michael Coleman ◽  
...  
Keyword(s):  
High Resolution ◽  
Light Sheet ◽  
High Resolution Imaging ◽  
Light Sheet Microscopy ◽  
Resolution Imaging ◽  
Living Organisms

scholarly journals Converting lateral scanning into axial focusing to speed up three-dimensional microscopy

2020 ◽  
Vol 9 (1) ◽  
Author(s):  
Tonmoy Chakraborty ◽  
Bingying Chen ◽  
Stephan Daetwyler ◽  
Bo-Jui Chang ◽  
Oliver Vanderpoorten ◽  
...  
Keyword(s):  
High Resolution ◽  
Spherical Aberration ◽  
Optical Design ◽  
Light Sheet ◽  
Three Dimensions ◽  
High Resolution Imaging ◽  
Light Sheet Microscopy ◽  
Scanning Rate ◽  
Resolution Imaging ◽  
Axial Scanning

Abstract In optical microscopy, the slow axial scanning rate of the objective or the sample has traditionally limited the speed of volumetric imaging. Recently, by conjugating either a movable mirror to the image plane in a remote-focusing geometry or an electrically tuneable lens (ETL) to the back focal plane, rapid axial scanning has been achieved. However, mechanical actuation of a mirror limits the axial scanning rate (usually only 10–100 Hz for piezoelectric or voice coil-based actuators), while ETLs introduce spherical and higher-order aberrations that prevent high-resolution imaging. In an effort to overcome these limitations, we introduce a novel optical design that transforms a lateral-scan motion into a spherical aberration-free axial scan that can be used for high-resolution imaging. Using a galvanometric mirror, we scan a laser beam laterally in a remote-focusing arm, which is then back-reflected from different heights of a mirror in the image space. We characterize the optical performance of this remote-focusing technique and use it to accelerate axially swept light-sheet microscopy by an order of magnitude, allowing the quantification of rapid vesicular dynamics in three dimensions. We also demonstrate resonant remote focusing at 12 kHz with a two-photon raster-scanning microscope, which allows rapid imaging of brain tissues and zebrafish cardiac dynamics with diffraction-limited resolution.

scholarly journals A Versatile Oblique Plane Microscope for Large-Scale and High-Resolution Imaging of Subcellular Dynamics

2020 ◽  
Author(s):  
Etai Sapoznik ◽  
Bo-Jui Chang ◽  
Jaewon Huh ◽  
Robert J. Ju ◽  
Evgenia V. Azarova ◽  
...  
Keyword(s):  
High Resolution ◽  
Numerical Aperture ◽  
Light Sheet ◽  
Membrane Dynamics ◽  
Field Of View ◽  
Large Field ◽  
Endoplasmic Reticulum Membrane ◽  
High Resolution Imaging ◽  
Light Sheet Microscopy ◽  
Resolution Imaging

AbstractWe present an Oblique Plane Microscope that uses a bespoke glass-tipped tertiary objective to improve the resolution, field of view, and usability over previous variants. Owing to its high numerical aperture optics, this microscope achieves lateral and axial resolutions that are comparable to the square illumination mode of Lattice Light-Sheet Microscopy, but in a user friendly and versatile format. Given this performance, we demonstrate high-resolution imaging of clathrin-mediated endocytosis, vimentin, the endoplasmic reticulum, membrane dynamics, and Natural Killer-mediated cytotoxicity. Furthermore, we image biological phenomena that would be otherwise challenging or impossible to perform in a traditional light-sheet microscope geometry, including cell migration through confined spaces within a microfluidic device, subcellular photoactivation of Rac1, diffusion of cytoplasmic rheological tracers at a volumetric rate of 14 Hz, and large field of view imaging of neurons, developing embryos, and centimeter-scale tissue sections.

scholarly journals A versatile oblique plane microscope for large-scale and high-resolution imaging of subcellular dynamics

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Etai Sapoznik ◽  
Bo-Jui Chang ◽  
Jaewon Huh ◽  
Robert J Ju ◽  
Evgenia V Azarova ◽  
...  
Keyword(s):  
High Resolution ◽  
Numerical Aperture ◽  
Light Sheet ◽  
Membrane Dynamics ◽  
Field Of View ◽  
Large Field ◽  
Endoplasmic Reticulum Membrane ◽  
High Resolution Imaging ◽  
Light Sheet Microscopy ◽  
Resolution Imaging

We present an oblique plane microscope (OPM) that uses a bespoke glass-tipped tertiary objective to improve the resolution, field of view, and usability over previous variants. Owing to its high numerical aperture optics, this microscope achieves lateral and axial resolutions that are comparable to the square illumination mode of lattice light-sheet microscopy, but in a user friendly and versatile format. Given this performance, we demonstrate high-resolution imaging of clathrin-mediated endocytosis, vimentin, the endoplasmic reticulum, membrane dynamics, and Natural Killer-mediated cytotoxicity. Furthermore, we image biological phenomena that would be otherwise challenging or impossible to perform in a traditional light-sheet microscope geometry, including cell migration through confined spaces within a microfluidic device, subcellular photoactivation of Rac1, diffusion of cytoplasmic rheological tracers at a volumetric rate of 14 Hz, and large field of view imaging of neurons, developing embryos, and centimeter-scale tissue sections.

LONG-TERM SAFETY, HIGH-RESOLUTION IMAGING, AND TISSUE TEMPERATURE MODELING OF SUBVISIBLE DIODE MICROPULSE PHOTOCOAGULATION FOR RETINOVASCULAR MACULAR EDEMA

Retina ◽  
2012 ◽  
Vol 32 (2) ◽  
pp. 375-386 ◽  
Author(s):  
Jeffrey K. Luttrull ◽  
Christopher Sramek ◽  
Daniel Palanker ◽  
Charles J. Spink ◽  
David C. Musch
Keyword(s):  
High Resolution ◽  
Macular Edema ◽  
Tissue Temperature ◽  
High Resolution Imaging ◽  
Temperature Modeling ◽  
Resolution Imaging

Long-Term, High-Resolution Imaging in the Neocortex In Vivo

2008 ◽  
Vol 2008 (2) ◽  
pp. pdb.prot4902-pdb.prot4902 ◽  
Author(s):  
B. E. Chen ◽  
J. T. Trachtenberg ◽  
A. J.G.D. Holtmaat ◽  
K. Svoboda
Keyword(s):  
High Resolution ◽  
High Resolution Imaging ◽  
Resolution Imaging

scholarly journals Light sheet theta microscopy for rapid high-resolution imaging of large biological samples

BMC Biology ◽  
2018 ◽  
Vol 16 (1) ◽  
Author(s):  
Bianca Migliori ◽  
Malika S. Datta ◽  
Christophe Dupre ◽  
Mehmet C. Apak ◽  
Shoh Asano ◽  
...  
Keyword(s):  
High Resolution ◽  
Biological Samples ◽  
Light Sheet ◽  
High Resolution Imaging ◽  
Resolution Imaging

Design and Validation of a Low-Cost Microscope for Diagnostics in the Developing World

2009 ◽  
Vol 17 (6) ◽  
pp. 16-19 ◽  
Author(s):  
B. Cline ◽  
R. Luo ◽  
K. Kuhlmann
Keyword(s):  
High Resolution ◽  
Infectious Diseases ◽  
Developing World ◽  
Low Cost ◽  
High Resolution Imaging ◽  
Microscope Examination ◽  
Resource Limited Settings ◽  
Resource Limited ◽  
Resolution Imaging

Many infectious diseases prevalent in the developing world, including malaria and tuberculosis, are difficult to diagnose on the basis of symptoms alone but can be accurately detected using microscope examination. Currently the expense, size, and fragility of optical microscopes impede their widespread use in resource-limited settings. Addressing these obstacles facing microscopy in the developing world is a pressing need; over 800,000 people, primarily children in Africa, die annually of malaria, and more than 1,500,000 people die annually of tuberculosis [1][2]. The aim of this study is to design and validate a microscope for use in the developing world that combines high-resolution imaging, extreme affordability, and long-term durability.

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