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.

scholarly journals Super-resolution nanoscopy by coherent control on nanoparticle emission

2020 ◽  
Vol 6 (16) ◽  
pp. eaaw6579
Author(s):  
Congyue Liu ◽  
Wei Liu ◽  
Shufeng Wang ◽  
Hongjia Li ◽  
Zhilong Lv ◽  
...  
Keyword(s):  
Coherent Control ◽  
High Efficiency ◽  
Super Resolution ◽  
General Strategy ◽  
Wide Field ◽  
Multiphoton Imaging ◽  
Two Photon ◽  
Characteristic Emission ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

Super-resolution nanoscopy based on wide-field microscopic imaging provided high efficiency but limited resolution. Here, we demonstrate a general strategy to push its resolution down to ~50 nm, which is close to the range of single molecular localization microscopy, without sacrificing the wide-field imaging advantage. It is done by actively and simultaneously modulating the characteristic emission of each individual emitter at high density. This method is based on the principle of excited state coherent control on single-particle two-photon fluorescence. In addition, the modulation efficiently suppresses the noise for imaging. The capability of the method is verified both in simulation and in experiments on ZnCdS quantum dot–labeled films and COS7 cells. The principle of coherent control is generally applicable to single-multiphoton imaging and various probes.

scholarly journals A technique for resolution assessment in blind-SIM experiments

2021 ◽  
Author(s):  
Imen Boujmil ◽  
Giancarlo Ruocco ◽  
Marco Leonetti
Keyword(s):  
High Resolution ◽  
Biological Sample ◽  
A Priori ◽  
Resolution Enhancement ◽  
Super Resolution ◽  
Experimental Setup ◽  
Wide Field ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Resolution Measurement

Super resolution techniques are an excellent alternative to wide field microscopy, providing high resolution also in (typically fragile) biological sample. Among the various super resolution techniques, Structured Illumination Microscopy (SIM) improve resolution by employing multiple illumination patterns to be deconvolved with a dedicated software. In the case of blind SIM techniques, unknown patterns, such as speckles, are used, thus providing super resolved images, nearly unaffected by aberrations with a simplified experimental setup. Scattering Assisted Imaging, a special blind SIM technique, exploits an illumination PSF (speckle grains size), smaller than the collection PSF (defined by the collection objectives), to surpass the typical SIM resolution enhancement. However, if SAI is used, it is very difficult to extract the resolution enhancement form a priori considerations. In this paper we propose a protocol and experimental setup for the resolution measurement, demonstrating the resolution enhancement for different collection PSF values.

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.

Diffusing Wave Spectroscopy: The Dynamics of Multiply Scattering Media

1991 ◽  
Vol 253 ◽  
Author(s):  
D. A. Weitz ◽  
D. J. Pine ◽  
D. J. Durian ◽  
J. X. Zhu
Keyword(s):  
Hydrodynamic Interaction ◽  
Colloidal Particles ◽  
Mean Free Path ◽  
Diffusing Wave Spectroscopy ◽  
Scattering Medium ◽  
Scattering Media ◽  
Physical Processes ◽  
Total Light ◽  
Path Lengths ◽  
Physical Phenomena

ABSTRACTWhen light is very strongly multiply scattered by a medium, its propagation can be well described by a diffusion approximation. This allows important, measurable quantities to be calculated theoretically and interpreted. Thus, for example the total light transmitted through a sample can be used to determine the transport mean free path which characterizes the diffusive transport of the light. In addition, the distribution of path lengths followed by the diffusing light can be determined. This distribution can in turn be used to interpret the temporal fluctuations of the scattered intensity that arise due to the motion of the scattering medium. Therefore, traditional quasielastic, or dynamic, light scattering can be extended to the strongly multiple scattering limit. This technique is called Diffusing Wave Spectroscopy (DWS), and allows useful information about the dynamics of the medium to be determined. Furthermore, new physical processes can be studiedusing DWS. For example, DWS is sensitive to very small motions of colloidal particles: motion of 1 pm diameter particles can be resolved on lengths of∼ 5 Å using light with a wavelength of 0.5 μm. New physical phenomena are probed when motion on these length scales is observed. In particular, the time evolution of the hydrodynamic interaction between concentrated colloidal particles can be resolved. In addition, DWScan also probe spatially rare events since the light paths sample a large volume of the sample. This allows DWS to probe very slow dynamics, making it useful for the study of materials such as foams. This talk reviews the fundamentals of DWS and highlights some ofits unique applications.

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 3D-resolved fluorescence and phosphorescence lifetime imaging using temporal focusing wide-field two-photon excitation

2012 ◽  
Vol 20 (24) ◽  
pp. 26219 ◽  
Author(s):  
Heejin Choi ◽  
Dimitrios S. Tzeranis ◽  
Jae Won Cha ◽  
Philippe Clémenceau ◽  
Sander J. G. de Jong ◽  
...  
Keyword(s):  
Wide Field ◽  
Phosphorescence Lifetime ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Photon Excitation ◽  
Temporal Focusing ◽  
Phosphorescence Lifetime Imaging ◽  
Fluorescence And Phosphorescence ◽  
Two Photon Excitation
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