scholarly journals Highly Sensitive Shack–Hartmann Wavefront Sensor: Application to Non-Transparent Tissue Mimic Imaging with Adaptive Light-Sheet Fluorescence Microscopy

2019 ◽  
Vol 2 (3) ◽  
pp. 59 ◽  
Author(s):  
Brajones ◽  
Clouvel ◽  
Dovillaire ◽  
Levecq ◽  
Lorenzo
Keyword(s):  
Adaptive Optics ◽  
Three Dimensional ◽  
High Sensitivity ◽  
Light Sheet ◽  
Wavefront Sensor ◽  
Spatial Frequencies ◽  
Living Tissues ◽  
Guide Star ◽  
Two Photon Fluorescence ◽  
Subcellular Resolution

High-quality in-depth imaging of three-dimensional samples remains a major challenge in modern microscopy. Selective plane illumination microscopy (SPIM) is a widely used technique that enables imaging of living tissues with subcellular resolution. However, scattering, absorption, and optical aberrations limit the depth at which useful imaging can be done. Adaptive optics (AOs) is a method capable of measuring and correcting aberrations in different kinds of fluorescence microscopes, thereby improving the performance of the optical system. We have incorporated a wavefront sensor adaptive optics scheme to SPIM (WAOSPIM) to correct aberrations induced by optically-thick samples, such as multi-cellular tumor spheroids (MCTS). Two-photon fluorescence provides us with a tool to produce a weak non-linear guide star (NGS) in any region of the field of view. The faintness of NGS; however, led us to develop a high-sensitivity Shack–Hartmann wavefront sensor (SHWS). This paper describes this newly developed SHWS and shows the correction capabilities of WAOSPIM using NGS in thick, inhomogeneous samples like MCTS. We report improvements of up to 79% for spatial frequencies corresponding to cellular and subcellular size features.

scholarly journals Full three-dimensional imaging deep through multicellular thick samples with subcellular resolution by structured illumination microscopy and adaptive optics

2020 ◽  
Author(s):  
Ruizhe Lin ◽  
Edward T. Kipreos ◽  
Jie Zhu ◽  
Chang Hyun Khang ◽  
Peter Kner
Keyword(s):  
Adaptive Optics ◽  
Three Dimensional ◽  
Structured Illumination ◽  
Axial Resolution ◽  
Structured Illumination Microscopy ◽  
Optical Aberrations ◽  
C Elegans ◽  
Culture Cells ◽  
Tissue Culture Cells ◽  
Subcellular Resolution

AbstractStructured Illumination Microscopy enables live imaging with resolutions of ~120 nm. Unfortunately, optical aberrations can lead to loss of resolution and artifacts in Structured Illumination Microscopy rendering the technique unusable in samples thicker than a single cell. Here we report on the combination of Adaptive Optics and Structured Illumination Microscopy enabling imaging with 140 nm lateral and 585 nm axial resolution in tissue culture cells, C. elegans, and rice blast fungus. We demonstrate that AO improves resolution and reduces artifacts, making full 3D SIM possible in thicker samples.

scholarly journals A telescope-ready approach for modal compensation of pyramid wavefront sensor optical gain

2019 ◽  
Vol 629 ◽  
pp. A107 ◽  
Author(s):  
V. Deo ◽  
É. Gendron ◽  
G. Rousset ◽  
F. Vidal ◽  
A. Sevin ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Optical Gain ◽  
High Sensitivity ◽  
Wavefront Sensor ◽  
Scientific Instrument ◽  
Nonlinearity Error ◽  
Operation Conditions ◽  
Gain Variation ◽  
Compensation Technique ◽  
Large Telescopes

The pyramid wavefront sensor (PWFS) is the currently preferred design for high-sensitivity adaptive optics (AO) systems for extremely large telescopes (ELTs). Yet, nonlinearities of the signal retrieved from the PWFS pose a significant problem for achieving the full correction potential using this sensor, a problem that will only worsen with the increasing dimension of telescopes. This paper investigates the so-called optical gain (OG) phenomenon, a sensitivity reduction and an overall modification of the sensor response induced by the residual wavefront itself, with considerable effects in standard observation conditions for ELT-sized AO systems. Through extensive numerical analysis, this work proposes a formalism to measure and minimize the first-order nonlinearity error caused by optical gain variation, which uses a modal compensation technique of the calibrated reconstructor; this enables a notable increase in performance in faint guide stars or important seeing scenarios, for example from 16 to 30% H-band Strehl ratio for a sixteenth magnitude star in r0 = 13 cm turbulence. Beyond the performance demonstrated by this compensation, a complete algorithm for realistic operation conditions is designed, which from dithering a few deformable mirror modes retrieves the optimal gains and updates the command matrix accordingly. The performance of this self-updating technique – which successfully allows automatic OG compensation regardless of the turbulent conditions, and its minimal interference with the scientific instrument are demonstrated through extensive end-to-end numerical simulations, all at the scale of an ELT instrument single-conjugate AO system.

scholarly journals Assessing and mitigating alignment defects of the pyramid wavefront sensor: a translation insensitive control method

2018 ◽  
Vol 619 ◽  
pp. A56 ◽  
Author(s):  
V. Deo ◽  
É. Gendron ◽  
G. Rousset ◽  
F. Vidal ◽  
T. Buey ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Control Method ◽  
Wavefront Sensor ◽  
Sampling Errors ◽  
Gradient Sensing ◽  
Design Factor ◽  
Performance Loss ◽  
Spatial Frequencies ◽  
Plane Wavefront ◽  
The Impact

The pyramid wavefront sensor (PWFS) is the currently preferred design for adaptive optics (AO) systems for extremely large telescopes, as focal plane wavefront sensing bears potential for a large intrinsic sensitivity gain when compared to Shack–Hartmann (SH) sensors. Yet, obtaining a high quality pyramidal prism and a model-consistent assembly remains a critical design factor. We demonstrate that the traditional gradient sensing controller is extremely sensitive to prism shape defects and assembly misalignments. We show that even optimal registration of quadrants on the detector may be insufficient to prevent misalignment induced performance loss through a theoretical analysis of the impact of detection plane quadrants sampling errors and individual translations, which may be induced by a variety of mechanical defects. These misalignments displace wavefront information to terms not included in the conventional gradient-like slopes maps and high spatial frequencies become invisible to the sole X− and Y− axis differences. We introduce expanded space control (ESC) for quad-cell signal by generalizing output measurements of the PWFS and demonstrate its insensitivity to misalignment-induced information loss, therefore dramatically relaxing machining and alignment constraints for PWFS engineering. This work presents the theoretical developments leading to ESC design, along with validating performance and robustness results, both in end-to-end numerical simulations and on a PWFS demonstrator bench at LESIA.

Accurate laser guide star wavefront sensor simulation for the E-ELT first light adaptive optics module

2016 ◽  
Author(s):  
Mauro Patti ◽  
Laura Schreiber ◽  
Carmelo Arcidiacono ◽  
Giovanni Bregoli ◽  
Paolo Ciliegi ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Wavefront Sensor ◽  
Laser Guide Star ◽  
Guide Star ◽  
Laser Guide

scholarly journals Imaging plant germline differentiation within Arabidopsis flowers by light sheet microscopy

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Sona Valuchova ◽  
Pavlina Mikulkova ◽  
Jana Pecinkova ◽  
Jana Klimova ◽  
Michal Krumnikl ◽  
...  
Keyword(s):  
Higher Plants ◽  
Three Dimensional ◽  
Cell Lineage ◽  
Light Sheet ◽  
Three Dimensional Model ◽  
Light Sheet Microscopy ◽  
Meiotic Chromosomes ◽  
Short Period ◽  
Germline Differentiation ◽  
Subcellular Resolution

In higher plants, germline differentiation occurs during a relatively short period within developing flowers. Understanding of the mechanisms that govern germline differentiation lags behind other plant developmental processes. This is largely because the germline is restricted to relatively few cells buried deep within floral tissues, which makes them difficult to study. To overcome this limitation, we have developed a methodology for live imaging of the germ cell lineage within floral organs of Arabidopsis using light sheet fluorescence microscopy. We have established reporter lines, cultivation conditions, and imaging protocols for high-resolution microscopy of developing flowers continuously for up to several days. We used multiview imagining to reconstruct a three-dimensional model of a flower at subcellular resolution. We demonstrate the power of this approach by capturing male and female meiosis, asymmetric pollen division, movement of meiotic chromosomes, and unusual restitution mitosis in tapetum cells. This method will enable new avenues of research into plant sexual reproduction.

scholarly journals Imaging plant germline differentiation within Arabidopsis flower by light sheet microscopy

2019 ◽  
Author(s):  
Sona Valuchova ◽  
Pavlina Mikulkova ◽  
Jana Pecinkova ◽  
Jana Klimova ◽  
Michal Krumnikl ◽  
...  
Keyword(s):  
Higher Plants ◽  
Three Dimensional ◽  
Cell Lineage ◽  
Light Sheet ◽  
Three Dimensional Model ◽  
Light Sheet Microscopy ◽  
Meiotic Chromosomes ◽  
Short Period ◽  
Germline Differentiation ◽  
Subcellular Resolution

AbstractIn higher plants, germline differentiation occurs during a relatively short period within developing flowers. Understanding of the mechanisms that govern germline differentiation lags behind other plant developmental processes. This is largely because the germline is restricted to relatively few cells buried deep within floral tissues, which makes them difficult to study. To overcome this limitation, we have developed a methodology for live imaging of the germ cell lineage within floral organs of Arabidopsis using light sheet fluorescence microscopy. We have established reporter lines, cultivation conditions, and imaging protocols for high-resolution microscopy of developing flowers continuously for up to several days. We used multiview imagining to reconstruct a three-dimensional model of a flower at subcellular resolution. We demonstrate the power of this approach by capturing male and female meiosis, asymmetric pollen division, movement of meiotic chromosomes, and unusual restitution mitosis in tapetum cells. This method will enable new avenues of research into plant sexual reproduction.

Dual-channel multiple natural guide star wavefront sensor for the E-ELT multiconjugate adaptive optics module

2012 ◽  
Author(s):  
Emiliano Diolaiti ◽  
Laura Schreiber ◽  
Italo Foppiani ◽  
Matteo Lombini
Keyword(s):  
Adaptive Optics ◽  
Wavefront Sensor ◽  
Dual Channel ◽  
Guide Star
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