scholarly journals High-resolution structural and functional deep brain imaging using adaptive optics three-photon microscopy

2021 ◽  
Author(s):  
Lina Streich ◽  
Juan Carlos Boffi ◽  
Ling Wang ◽  
Khaleel Alhalaseh ◽  
Matteo Barbieri ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Multiphoton Microscopy ◽  
Motion Artifacts ◽  
Tissue Imaging ◽  
Mammalian Brain ◽  
Neural Cells ◽  
Photon Excitation ◽  
Deep Tissue Imaging ◽  
Fibrous Astrocytes ◽  
And Function

AbstractMultiphoton microscopy has become a powerful tool with which to visualize the morphology and function of neural cells and circuits in the intact mammalian brain. However, tissue scattering, optical aberrations and motion artifacts degrade the imaging performance at depth. Here we describe a minimally invasive intravital imaging methodology based on three-photon excitation, indirect adaptive optics (AO) and active electrocardiogram gating to advance deep-tissue imaging. Our modal-based, sensorless AO approach is robust to low signal-to-noise ratios as commonly encountered in deep scattering tissues such as the mouse brain, and permits AO correction over large axial fields of view. We demonstrate near-diffraction-limited imaging of deep cortical spines and (sub)cortical dendrites up to a depth of 1.4 mm (the edge of the mouse CA1 hippocampus). In addition, we show applications to deep-layer calcium imaging of astrocytes, including fibrous astrocytes that reside in the highly scattering corpus callosum.

scholarly journals High-resolution structural and functional deep brain imaging using adaptive optics three-photon microscopy

2021 ◽  
Author(s):  
Lina Streich ◽  
Juan Boffi ◽  
Ling Wang ◽  
Khaleel Alhalaseh ◽  
Matteo Barbieri ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Motion Artifacts ◽  
Mammalian Brain ◽  
Neural Cells ◽  
Optical Aberrations ◽  
Photon Excitation ◽  
Tissue Scattering ◽  
Imaging Methodology ◽  
And Function

Multi-photon microscopy has become a powerful tool to visualize the morphology and function of neural cells and circuits in the intact mammalian brain. Yet, tissue scattering, optical aberrations, and motion artifacts degrade the achievable image quality with depth. Here we developed a minimally invasive intravital imaging methodology by combining three-photon excitation, indirect adaptive optics correction, and active electrocardiogram gating to achieve near-diffraction limited resolution up to a depth of 1.2mm in the mouse brain. We demonstrate near-diffraction-limited imaging of deep cortical and sub-cortical dendrites and spines as well as of calcium transients in deep-layer astrocytes in vivo.

scholarly journals Improving the Way We See: Adaptive Optics Based Optical Microscopy for Deep-Tissue Imaging

2021 ◽  
Vol 9 ◽  
Author(s):  
Pranoy Sahu ◽  
Nirmal Mazumder
Keyword(s):  
Adaptive Optics ◽  
Cellular Level ◽  
Optical Methods ◽  
Tissue Imaging ◽  
Excitation Beam ◽  
Recent Developments ◽  
Deep Tissue Imaging ◽  
And Function ◽  
Tools And Techniques ◽  
Two Photon Fluorescence

With the recent developments in optical imaging tools and techniques, scientists are now able to image deeper regions of the tissue with greater resolution and accuracy. However, light scattering while imaging deeper regions of a biological tissue remains a fundamental issue. Presence of lipids, proteins and nucleic acids in the tissue makes it inhomogeneous for a given wavelength of light. Two-photon fluorescence (TPF) microscopy supplemented with improved invasive optical tools allows functional imaging in awake behaving mammals in an unprecedented manner. Similarly, improved optical methods conjugated with previously existing scanning laser ophthalmoscopy (SLO) has paved diffraction-limited retinal imaging. With the evolving technology, scientists are now able to resolve biological structures and function at the sub-cellular level. Wavefront correcting methods like adaptive optics (AO) has been implemented in correcting tissue or optical-based distortions, shaping the excitation beam in 3D-holography to target multiple neurons. And more recently, AO-based SLO is implemented for eye imaging both in research and clinical settings. In this review, we discuss some of the recent improvements in TPF microscopy with the application of AO for wavefront corrections and its recent application in brain imaging as well as ophthalmoscopy.

Infrared multiphoton microscopy: subcellular-resolved deep tissue imaging

2009 ◽  
Vol 20 (1) ◽  
pp. 54-62 ◽  
Author(s):  
Volker Andresen ◽  
Stephanie Alexander ◽  
Wolfgang-Moritz Heupel ◽  
Markus Hirschberg ◽  
Robert M. Hoffman ◽  
...  
Keyword(s):  
Multiphoton Microscopy ◽  
Tissue Imaging ◽  
Deep Tissue ◽  
Deep Tissue Imaging

Lysosome-targetable polythiophene nanoparticles for two-photon excitation photodynamic therapy and deep tissue imaging

2017 ◽  
Vol 5 (20) ◽  
pp. 3651-3657 ◽  
Author(s):  
Shaojing Zhao ◽  
Guangle Niu ◽  
Feng Wu ◽  
Li Yan ◽  
Hongyan Zhang ◽  
...  
Keyword(s):  
Photodynamic Therapy ◽  
Quantum Yield ◽  
Fluorescence Imaging ◽  
Cross Section ◽  
Tissue Imaging ◽  
Deep Tissue ◽  
Two Photon ◽  
Photon Excitation ◽  
Deep Tissue Imaging ◽  
Two Photon Excitation

Polythiophene nanoparticles with large TPA cross section and high1O2generation quantum yield have been developed for simultaneous lysosome-targetable fluorescence imaging and photodynamic therapy.

Multiple guide stars optimization in conjugate adaptive optics for deep tissue imaging

2020 ◽  
Vol 459 ◽  
pp. 124891
Author(s):  
Chenxue Wu ◽  
Jiajia Chen ◽  
Biwei Zhang ◽  
Yao Zheng ◽  
Xinpei Zhu ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Tissue Imaging ◽  
Deep Tissue ◽  
Deep Tissue Imaging

scholarly journals Deep Learning Assisted Zonal Adaptive Aberration Correction

2021 ◽  
Vol 8 ◽  
Author(s):  
Biwei Zhang ◽  
Jiazhu Zhu ◽  
Ke Si ◽  
Wei Gong
Keyword(s):  
Deep Learning ◽  
Adaptive Optics ◽  
Degrees Of Freedom ◽  
Correction Method ◽  
Tissue Imaging ◽  
Optical Aberrations ◽  
Volume Imaging ◽  
Adaptive Correction ◽  
Average Mean Square Error ◽  
Deep Tissue Imaging

Deep learning (DL) has been recently applied to adaptive optics (AO) to correct optical aberrations rapidly in biomedical imaging. Here we propose a DL assisted zonal adaptive correction method to perform corrections of high degrees of freedom while maintaining the fast speed. With a trained DL neural network, the pattern on the correction device which is divided into multiple zone phase elements can be directly inferred from the aberration distorted point-spread function image in this method. The inference can be completed in 12.6 ms with the average mean square error 0.88 when 224 zones are used. The results show a good performance on aberrations of different complexities. Since no extra device is required, this method has potentials in deep tissue imaging and large volume imaging.

Signal to Noise Considerations in Single Pixel Multiphoton Microscopy

2020 ◽  
Vol 5 (5) ◽  
pp. 1-2
Author(s):  
Dushan Wadduwage
Keyword(s):  
Multiphoton Microscopy ◽  
Tissue Imaging ◽  
Wide Field ◽  
Signal To Noise ◽  
Deep Tissue ◽  
Multiplexed Imaging ◽  
Deep Tissue Imaging ◽  
Single Pixel ◽  
Noise Metrics

Single-pixel imaging geometries for wide-field multiphoton microscopy (SPx-MPM) have emerged as a contender to conventional point-scanning multiphoton systems (PS-MPM) for deep tissue imaging. These systems are thought to be faster due to their multiplexed imaging capabilities with higher photon throughput. In this study we numerically compare the signal to noise metrics of the SPx-MPM to the PS-MPM systems. Our results suggest that PS-MPM systems outperform SPx-MPM systems, despite their higher photon throughput.

scholarly journals Overcoming the penetration depth limit in optical microscopy: Adaptive optics and wavefront shaping

2019 ◽  
Vol 12 (04) ◽  
pp. 1930002 ◽  
Author(s):  
Cheolwoo Ahn ◽  
Byungjae Hwang ◽  
Kibum Nam ◽  
Hyungwon Jin ◽  
Taeseong Woo ◽  
...  
Keyword(s):  
High Resolution ◽  
Optical Microscopy ◽  
Adaptive Optics ◽  
Single Cells ◽  
Tissue Imaging ◽  
Future Research ◽  
Deep Tissue ◽  
Depth Limit ◽  
Deep Tissue Imaging ◽  
Wavefront Shaping

Despite the unique advantages of optical microscopy for molecular specific high resolution imaging of living structure in both space and time, current applications are mostly limited to research settings. This is due to the aberrations and multiple scattering that is induced by the inhomogeneous refractive boundaries that are inherent to biological systems. However, recent developments in adaptive optics and wavefront shaping have shown that high resolution optical imaging is not fundamentally limited only to the observation of single cells, but can be significantly enhanced to realize deep tissue imaging. To provide insight into how these two closely related fields can expand the limits of bio imaging, we review the recent progresses in their performance and applicable range of studies as well as potential future research directions to push the limits of deep tissue imaging.

scholarly journals Cross-species Transcriptomic Comparison of In Vitro and In Vivo Mammalian Neural Cells

2015 ◽  
Vol 9 ◽  
pp. BBI.S33124 ◽  
Author(s):  
Peter R. LoVerso ◽  
Christopher M. Wachter ◽  
Feng Cui
Keyword(s):  
Gene Expression ◽  
Transcript Abundance ◽  
Cell Types ◽  
Mammalian Brain ◽  
Neural Cells ◽  
Rna Seq ◽  
Commercial Sources ◽  
And Function

The mammalian brain is characterized by distinct classes of cells that differ in morphology, structure, signaling, and function. Dysregulation of gene expression in these cell populations leads to various neurological disorders. Neural cells often need to be acutely purified from animal brains for research, which requires complicated procedure and specific expertise. Primary culture of these cells in vitro is a viable alternative, but the differences in gene expression of cells grown in vitro and in vivo remain unclear. Here, we cultured three major neural cell classes of rat brain (ie, neurons, astrocytes, and oligodendrocyte precursor cells [OPCs]) obtained from commercial sources. We measured transcript abundance of these cell types by RNA sequencing (RNA-seq) and compared with their counterparts acutely purified from mouse brains. Cross-species RNA-seq data analysis revealed hundreds of genes that are differentially expressed between the cultured and acutely purified cells. Astrocytes have more such genes compared to neurons and OPCs, indicating that signaling pathways are greatly perturbed in cultured astrocytes. This dataset provides a powerful resource to demonstrate the similarities and differences of biological processes in mammalian neural cells grown in vitro and in vivo at the molecular level.

scholarly journals Ultrasound Imaging of Gene Expression in Mammalian Cells

2019 ◽  
Author(s):  
Arash Farhadi ◽  
Gabrielle H. Ho ◽  
Daniel P. Sawyer ◽  
Raymond W. Bourdeau ◽  
Mikhail G. Shapiro
Keyword(s):  
Gene Expression ◽  
Mammalian Cells ◽  
Reporter Genes ◽  
Tissue Imaging ◽  
Cellular Functions ◽  
Cellular Processes ◽  
Deep Tissue Imaging ◽  
And Function ◽  
Expression In Mammalian Cells

ABSTRACTThe study of cellular processes occurring inside intact organisms and the development of cell-based diagnostic and therapeutic agents requires methods to visualize cellular functions such as gene expression in deep tissues. Ultrasound is a widely used biomedical technology enabling deep-tissue imaging with high spatial and temporal resolution. However, no genetically encoded molecular reporters are available to connect ultrasound contrast to gene expression in mammalian cells. To address this limitation, we introduce the first mammalian acoustic reporter genes. Starting with an eleven-gene polycistronic gene cluster derived from bacteria, we engineered a eukaryotic genetic program whose introduction into mammalian cells results in the expression of a unique class of intracellular air-filled protein nanostructures called gas vesicles. The scattering of ultrasound by these nanostructures allows mammalian cells to be visualized at volumetric densities below 0.5%, enables the monitoring of dynamic circuit-driven gene expression, and permits high-resolution imaging of gene expression in living animals. These mammalian acoustic reporter genes enable previously impossible approaches to monitoring the location, viability and function of mammalian cellsin vivo.

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