scholarly journals Fast widefield imaging of neuronal structure and function with optical sectioning in vivo

2020 ◽  
Vol 6 (19) ◽  
pp. eaaz3870 ◽  
Author(s):  
Ziwei Li ◽  
Qinrong Zhang ◽  
Shih-Wei Chou ◽  
Zachary Newman ◽  
Raphaël Turcotte ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
In Vivo Imaging ◽  
Neuromuscular Junctions ◽  
Structure And Function ◽  
Structured Illumination ◽  
Neuronal Structure ◽  
Optical Sectioning ◽  
Structured Illumination Microscopy ◽  
And Function

Optical microscopy, owing to its noninvasiveness and subcellular resolution, enables in vivo visualization of neuronal structure and function in the physiological context. Optical-sectioning structured illumination microscopy (OS-SIM) is a widefield fluorescence imaging technique that uses structured illumination patterns to encode in-focus structures and optically sections 3D samples. However, its application to in vivo imaging has been limited. In this study, we optimized OS-SIM for in vivo neural imaging. We modified OS-SIM reconstruction algorithms to improve signal-to-noise ratio and correct motion-induced artifacts in live samples. Incorporating an adaptive optics (AO) module to OS-SIM, we found that correcting sample-induced optical aberrations was essential for achieving accurate structural and functional characterizations in vivo. With AO OS-SIM, we demonstrated fast, high-resolution in vivo imaging with optical sectioning for structural imaging of mouse cortical neurons and zebrafish larval motor neurons, and functional imaging of quantal synaptic transmission at Drosophila larval neuromuscular junctions.

Functional biomedical hydrogels for in vivo imaging

2016 ◽  
Vol 4 (48) ◽  
pp. 7793-7812 ◽  
Author(s):  
Kewen Lei ◽  
Qian Ma ◽  
Lin Yu ◽  
Jiandong Ding
Keyword(s):  
Real Time ◽  
In Vivo Imaging ◽  
Structure And Function ◽  
Non Invasive ◽  
The Status ◽  
And Function

In vivo imaging of biomedical hydrogels enables real-time and non-invasive visualization of the status of structure and function of hydrogels.

scholarly journals Differential structured illumination microendoscopy for in vivo imaging of molecular contrast agents

2016 ◽  
Vol 113 (39) ◽  
pp. 10769-10773 ◽  
Author(s):  
Pelham Keahey ◽  
Preetha Ramalingam ◽  
Kathleen Schmeler ◽  
Rebecca R. Richards-Kortum
Keyword(s):  
Contrast Agents ◽  
In Vivo Imaging ◽  
Fiber Optic ◽  
Adenocarcinoma In Situ ◽  
Structured Illumination ◽  
Optical Sectioning ◽  
Optical Elements ◽  
Fiber Optic System ◽  
Molecular Contrast

Fiber optic microendoscopy has shown promise for visualization of molecular contrast agents used to study disease in vivo. However, fiber optic microendoscopes have limited optical sectioning capability, and image contrast is limited by out-of-focus light generated in highly scattering tissue. Optical sectioning techniques have been used in microendoscopes to remove out-of-focus light but reduce imaging speed or rely on bulky optical elements that prevent in vivo imaging. Here, we present differential structured illumination microendoscopy (DSIMe), a fiber optic system that can perform structured illumination in real time for optical sectioning without any opto-mechanical components attached to the distal tip of the fiber bundle. We demonstrate the use of DSIMe during in vivo fluorescence imaging in patients undergoing surgery for cervical adenocarcinoma in situ. Images acquired using DSIMe show greater contrast than standard microendoscopy, improving the ability to detect cellular atypia associated with neoplasia.

scholarly journals Filopodome mapping identifies p130Cas as a mechanosensitive regulator of filopodia stability

2018 ◽  
Author(s):  
Guillaume Jacquemet ◽  
Rafael Saup ◽  
Hellyeh Hamidi ◽  
Mitro Miihkinen ◽  
Johanna Ivaska
Keyword(s):  
Binding Sites ◽  
Focal Adhesions ◽  
Structure And Function ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Cell Adhesion And Migration ◽  
And Migration ◽  
Cellular Behaviour ◽  
And Function ◽  
Fine Tune

AbstractFilopodia are adhesive cellular protrusions specialised in the detection of extracellular matrix (ECM)-derived cues. While ECM engagement at focal adhesions is known to trigger the recruitment of hundreds of proteins (“adhesome”) to fine-tune cellular behaviour, the components of the filopodia adhesions remain undefined. Here, we performed a structured illumination microscopy-based screen to map the localisation of 80 target proteins, linked to cell adhesion and migration, within filopodia. We demonstrate preferential enrichment of several adhesion proteins to either filopodia tips, filopodia shafts, or shaft subdomains suggesting divergent, spatially restricted functions for these proteins. Moreover, proteins with phospho-inositide (PI) binding sites are particularly enriched in filopodia. This, together with the strong localisation of PI(3,4)P2 in filopodia tips, predicts critical roles for PIs in regulating filopodia ultra-structure and function. Our mapping further reveals that filopodia adhesions consist of a unique set of proteins, the filopodome, that are distinct from classical nascent adhesions, focal adhesions and fibrillar adhesions. Using live imaging, we observe that filopodia adhesions can give rise to nascent adhesions, which, in turn, form focal adhesions. Finally, we demonstrate that p130Cas (BCAR1) is recruited to filopodia tips via its CCHD domain and acts as a mechanosensitive regulator of filopodia stability.

scholarly journals Dynamic super-resolution structured illumination imaging in the living brain

2019 ◽  
Vol 116 (19) ◽  
pp. 9586-9591 ◽  
Author(s):  
Raphaël Turcotte ◽  
Yajie Liang ◽  
Masashi Tanimoto ◽  
Qinrong Zhang ◽  
Ziwei Li ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Super Resolution ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Optical Aberrations ◽  
Sample Motion ◽  
Heterogeneous Tissue ◽  
Conventional Microscopy ◽  
Super Resolution Microscopy

Cells in the brain act as components of extended networks. Therefore, to understand neurobiological processes in a physiological context, it is essential to study them in vivo. Super-resolution microscopy has spatial resolution beyond the diffraction limit, thus promising to provide structural and functional insights that are not accessible with conventional microscopy. However, to apply it to in vivo brain imaging, we must address the challenges of 3D imaging in an optically heterogeneous tissue that is constantly in motion. We optimized image acquisition and reconstruction to combat sample motion and applied adaptive optics to correcting sample-induced optical aberrations in super-resolution structured illumination microscopy (SIM) in vivo. We imaged the brains of live zebrafish larvae and mice and observed the dynamics of dendrites and dendritic spines at nanoscale resolution.

Applications of phosphorescent materials for in-vivo imaging of brain structure and function

2016 ◽  
Author(s):  
Gregory Boverman ◽  
Xiaolei Shi ◽  
Victoria E. Cotero ◽  
Robert J. Filkins ◽  
Alok M. Srivastava ◽  
...  
Keyword(s):  
Brain Structure ◽  
In Vivo Imaging ◽  
Structure And Function ◽  
Brain Structure And Function ◽  
And Function

scholarly journals Reflective mirror-based line-scan adaptive optics OCT

2021 ◽  
Author(s):  
Vimal Prabhu Pandiyan ◽  
Xiaoyun Jiang ◽  
James A Kuchenbecker ◽  
Ramkumar Sabesan
Keyword(s):  
High Resolution ◽  
Adaptive Optics ◽  
Ex Vivo ◽  
Optical Design ◽  
Structure And Function ◽  
Line Scan ◽  
Reflective Mirror ◽  
Retinal Structure ◽  
And Function

Line-scan OCT, incorporated with adaptive optics (AO), offers high resolution, speed and sensitivity for imaging retinal structure and function in vivo. Here, we introduce its implementation with reflective mirror-based afocal telescopes, optimized for imaging light-induced retinal activity (optoretinography) and weak retinal reflections at the cellular scale. A non-planar optical design was followed based on previous recommendations with key differences specific to a line-scan geometry. The three beam paths fundamental to an OCT system -illumination/sample, detection, and reference - were modeled in Zemax optical design software to yield theoretically diffraction-limited performance over a 2.2 deg. field-of-view and 1.5 D vergence range at the eye's pupil. The performance for imaging retinal structure was exemplified by cellular-scale visualization of retinal ganglion cells, macrophages, foveal cones, and rods in human observers. The performance for functional imaging was exemplified by resolving the light-evoked optical changes in foveal cone photoreceptors where the spatial resolution was sufficient for cone spectral classification at an eccentricity 0.3 deg. from the foveal center. This enabled the first in vivo demonstration of reduced S-cone (short-wavelength cone) density in the human foveola, thus far observed only in ex vivo histological preparations. Together, the feasibility for high resolution imaging of retinal structure and function demonstrated here holds significant potential for basic science and translational applications.

scholarly journals In vivo imaging of photoreceptor structure and function in a non-human primate model of retinal degeneration

2017 ◽  
Vol 17 (7) ◽  
pp. 53
Author(s):  
Sarah Walters ◽  
Christina Schwarz ◽  
Robin Sharma ◽  
William S. Fischer ◽  
David DiLoreto ◽  
...  
Keyword(s):  
Retinal Degeneration ◽  
In Vivo Imaging ◽  
Structure And Function ◽  
Primate Model ◽  
And Function ◽  
Human Primate

scholarly journals Requirement of the Cep57-Cep63 Interaction for Proper Cep152 Recruitment and Centriole Duplication

2020 ◽  
Vol 40 (10) ◽  
Author(s):  
Zhuang Wei ◽  
Tae-Sung Kim ◽  
Jong Il Ahn ◽  
Lingjun Meng ◽  
Yaozong Chen ◽  
...  
Keyword(s):  
Self Assembly ◽  
Three Dimensional ◽  
Structure And Function ◽  
Binding Domain ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Centriole Duplication ◽  
And Function ◽  
Mutant Cells

ABSTRACT Cep57 has been characterized as a component of a pericentriolar complex containing Cep63 and Cep152. Interestingly, Cep63 and Cep152 self-assemble into a pericentriolar cylindrical architecture, and this event is critical for the orderly recruitment of Plk4, a key regulator of centriole duplication. However, the way in which Cep57 interacts with the Cep63-Cep152 complex and contributes to the structure and function of Cep63-Cep152 self-assembly remains unknown. We demonstrate that Cep57 interacts with Cep63 through N-terminal motifs and associates with Cep152 via Cep63. Three-dimensional structured illumination microscopy (3D-SIM) analyses suggested that the Cep57-Cep63-Cep152 complex is concentrically arranged around a centriole in a Cep57-in and Cep152-out manner. Cep57 mutant cells defective in Cep63 binding exhibited improper Cep63 and Cep152 localization and impaired Sas6 recruitment for procentriole assembly, proving the significance of the Cep57-Cep63 interaction. Intriguingly, Cep63 fused to a microtubule (MT)-binding domain of Cep57 functioned in concert with Cep152 to assemble around stabilized MTs in vitro. Thus, Cep57 plays a key role in architecting the Cep63-Cep152 assembly around centriolar MTs and promoting centriole biogenesis. This study may offer a platform to investigate how the organization and function of the pericentriolar architecture are altered by disease-associated mutations found in the Cep57-Cep63-Cep152 complex.

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