scholarly journals Wide-field multiphoton imaging of cellular dynamics in thick tissue by temporal focusing and patterned illumination

2011 ◽  
Vol 2 (3) ◽  
pp. 696 ◽  
Author(s):  
O. D. Therrien ◽  
B. Aubé ◽  
S. Pagès ◽  
P. De Koninck ◽  
D. Côté
Keyword(s):  
Wide Field ◽  
Cellular Dynamics ◽  
Multiphoton Imaging ◽  
Temporal Focusing ◽  
Patterned Illumination

scholarly journals Wide-field multiphoton imaging through scattering media without correction

2018 ◽  
Vol 4 (10) ◽  
pp. eaau1338 ◽  
Author(s):  
Adrià Escobet-Montalbán ◽  
Roman Spesyvtsev ◽  
Mingzhou Chen ◽  
Wardiya Afshar Saber ◽  
Melissa Andrews ◽  
...  
Keyword(s):  
A Priori ◽  
Super Resolution ◽  
Mean Free Path ◽  
Wide Field ◽  
Biomedical Sciences ◽  
Scattering Media ◽  
Multiphoton Imaging ◽  
Two Photon ◽  
Temporal Focusing ◽  
Path Lengths

Optical approaches to fluorescent, spectroscopic, and morphological imaging have made exceptional advances in the last decade. Super-resolution imaging and wide-field multiphoton imaging are now underpinning major advances across the biomedical sciences. While the advances have been startling, the key unmet challenge to date in all forms of optical imaging is to penetrate deeper. A number of schemes implement aberration correction or the use of complex photonics to address this need. In contrast, we approach this challenge by implementing a scheme that requires no a priori information about the medium nor its properties. Exploiting temporal focusing and single-pixel detection in our innovative scheme, we obtain wide-field two-photon images through various turbid media including a scattering phantom and tissue reaching a depth of up to seven scattering mean free path lengths. Our results show that it competes favorably with standard point-scanning two-photon imaging, with up to a fivefold improvement in signal-to-background ratio while showing significantly lower photobleaching.

Studies on wide-field-of-view multiphoton imaging using the flexible clinical multiphoton tomograph MPTflex

2012 ◽  
Author(s):  
Martin Weinigel ◽  
Hans Georg Breunig ◽  
Peter Fischer ◽  
Marcel Kellner-Hoefer ◽  
Rainer Bückle ◽  
...  
Keyword(s):  
Field Of View ◽  
Wide Field ◽  
Multiphoton Imaging ◽  
Wide Field Of View

scholarly journals Video-rate multimodal multiphoton imaging and three-dimensional characterization of cellular dynamics in wounded skin

2020 ◽  
Vol 13 (02) ◽  
pp. 2050007
Author(s):  
Joanne Li ◽  
Madison N. Wilson ◽  
Andrew J. Bower ◽  
Marina Marjanovic ◽  
Eric J. Chaney ◽  
...  
Keyword(s):  
Cell Tracking ◽  
Skin Diseases ◽  
Ex Vivo ◽  
Three Dimensional ◽  
Biological Information ◽  
Cellular Dynamics ◽  
Multiphoton Imaging ◽  
Cellular Events ◽  
Analysis Platform

To date, numerous studies have been performed to elucidate the complex cellular dynamics in skin diseases, but few have attempted to characterize these cellular events under conditions similar to the native environment. To address this challenge, a three-dimensional (3D) multimodal analysis platform was developed for characterizing in vivo cellular dynamics in skin, which was then utilized to process in vivo wound healing data to demonstrate its applicability. Special attention is focused on in vivo biological parameters that are difficult to study with ex vivo analysis, including 3D cell tracking and techniques to connect biological information obtained from different imaging modalities. These results here open new possibilities for evaluating 3D cellular dynamics in vivo, and can potentially provide new tools for characterizing the skin microenvironment and pathologies in the future.

scholarly journals Wide-field three-dimensional optical imaging using temporal focusing for holographically trapped microparticles

2015 ◽  
Vol 40 (21) ◽  
pp. 4847 ◽  
Author(s):  
Roman Spesyvtsev ◽  
Helen A. Rendall ◽  
Kishan Dholakia
Keyword(s):  
Optical Imaging ◽  
Three Dimensional ◽  
Wide Field ◽  
Temporal Focusing

scholarly journals De-scattering with Excitation Patterning enables rapid wide-field imaging through scattering media

2021 ◽  
Vol 7 (28) ◽  
pp. eaay5496
Author(s):  
Cheng Zheng ◽  
Jong Kang Park ◽  
Murat Yildirim ◽  
Josiah R. Boivin ◽  
Yi Xue ◽  
...  
Keyword(s):  
High Resolution ◽  
Structural Features ◽  
Computational Imaging ◽  
Tissue Imaging ◽  
Wide Field ◽  
Scattering Media ◽  
Temporal Focusing ◽  
Field Imaging ◽  
Wide Field Imaging

Nonlinear optical microscopy has enabled in vivo deep tissue imaging on the millimeter scale. A key unmet challenge is its limited throughput especially compared to rapid wide-field modalities that are used ubiquitously in thin specimens. Wide-field imaging methods in tissue specimens have found successes in optically cleared tissues and at shallower depths, but the scattering of emission photons in thick turbid samples severely degrades image quality at the camera. To address this challenge, we introduce a novel technique called De-scattering with Excitation Patterning or “DEEP,” which uses patterned nonlinear excitation followed by computational imaging–assisted wide-field detection. Multiphoton temporal focusing allows high-resolution excitation patterns to be projected deep inside specimen at multiple scattering lengths due to the use of long wavelength light. Computational reconstruction allows high-resolution structural features to be reconstructed from tens to hundreds of DEEP images instead of millions of point-scanning measurements.

scholarly journals Two-photon patterned photostimulation with low-power, high-efficiency and reliable single-cell optogenetic control

2022 ◽  
Author(s):  
Yifan Wang ◽  
Yao Zheng ◽  
Yongxian Xu ◽  
Rongrong Li ◽  
Yameng Zheng ◽  
...  
Keyword(s):  
Low Power ◽  
Single Cell ◽  
Laser Power ◽  
High Efficiency ◽  
Excitation Density ◽  
Two Photon ◽  
Photon Excitation ◽  
Temporal Focusing ◽  
Patterned Illumination ◽  
Two Photon Excitation

Two-photon optogenetics enables selectively stimulating individual cells for manipulating neuronal ensembles. As the general photostimulation strategy, the patterned two-photon excitation has enabled millisecond-timescale activation for single or multiple neurons, but its activation efficiency is suffered from high laser power due to low beam-modulation efficiency. Here, we develop a high-efficiency beam-shaping method based on the Gerchberg-Saxton (GS) algorithm with spherical-distribution initial phase (GSSIP) to reduce the patterned two-photon excitation speckles and intensity. It can well control the phase of shaped beams to attain speckle-free accurate patterned illumination with an improvement of 44.21% in the modulation efficiency compared with that of the traditional GS algorithm. A combination of temporal focusing and the GSSIP algorithm (TF-GSSIP) achieves patterned focusing through 500-μm-thickness mouse brain slices, which is 2.5 times deeper than the penetration depth of TF-GS with the same signal-to-noise ratio (SNR). With our method, the laser power can be reduced to only 55.56% of that with traditional method (the temporal focusing with GS, TF-GS) to reliably evoke GCaMP6s response in C1V1-expressing cultured neurons with single-cell resolution. Besides, the photostimulation efficiency is remarkably increased by 80.19% at the same excitation density of 0.27 mW/μm2. This two-photon stimulation method with low-power, reliable and patterned illumination may pave the way for analyzing neural circuits and neural coding and decoding mechanism.

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