scholarly journals Three-dimensional cardiac architecture determined by two-photon microtomy

2009 ◽  
Vol 14 (4) ◽  
pp. 044029 ◽  
Author(s):  
Hayden Huang ◽  
Catherine MacGillivray ◽  
Hyuk-Sang Kwon ◽  
Jan Lammerding ◽  
Jeffrey Robbins ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Two Photon

scholarly journals High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiang Lan Fan ◽  
Jose A. Rivera ◽  
Wei Sun ◽  
John Peterson ◽  
Henry Haeberle ◽  
...  
Keyword(s):  
Blood Flow ◽  
High Speed ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Fluorescence Signal ◽  
Imaging Method ◽  
Spatiotemporal Resolution ◽  
Two Photon

AbstractUnderstanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.

Two-Photon Excitation Imaging of Glucose Metabolism in Living Tissue

1997 ◽  
Vol 3 (S2) ◽  
pp. 305-306
Author(s):  
David W. Piston
Keyword(s):  
Confocal Microscopy ◽  
Collection Efficiency ◽  
Three Dimensional ◽  
Spatial Filter ◽  
Input Power ◽  
Photon Absorption ◽  
Focal Volume ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

Two-photon excitation microscopy (TPEM) provides attractive advantages over confocal microscopy for three-dimensionally resolved fluorescence imaging and photochemistry. It provides three-dimensional resolution and eliminates background equivalent to an ideal confocal microscope without requiring a confocal spatial filter, whose absence enhances fluorescence collection efficiency. This results in inherent submicron optical sectioning by excitation alone. In practice, TPEM is made possible by the very high local instantaneous intensity provided by a combination of diffraction-limited focusing of a single laser beam in the microscope and the temporal concentration of 100 femtosecond pulses generated by a mode-locked laser. Resultant peak excitation intensities are 106 times greater than the CW intensities used in confocal microscopy, but the pulse duty cycle of 10−5 limits the average input power to less than 10 mW, only slightly greater than the power normally used in confocal microscopy. Because of the intensity-squared dependence of the two-photon absorption, the excitation is limited to the focal volume.

scholarly journals Advanced Micro-Actuator/Robot Fabrication Using Ultrafast Laser Direct Writing and Its Remote Control

2020 ◽  
Vol 10 (23) ◽  
pp. 8563
Author(s):  
Sangmo Koo
Keyword(s):  
Three Dimensional ◽  
Processing Technique ◽  
Direct Writing ◽  
Laser Direct Writing ◽  
Precise Control ◽  
Two Photon ◽  
Non Invasive ◽  
Acoustic Control ◽  
Fs Laser ◽  
Invasive Control

Two-photon polymerization (TPP) based on the femtosecond laser (fs laser) direct writing technique in the realization of high-resolution three-dimensional (3D) shapes is spotlighted as a unique and promising processing technique. It is also interesting that TPP can be applied to various applications in not only optics, chemistry, physics, biomedical engineering, and microfluidics but also micro-robotics systems. Effort has been made to design innovative microscale actuators, and research on how to remotely manipulate actuators is also constantly being conducted. Various manipulation methods have been devised including the magnetic, optical, and acoustic control of microscale actuators, demonstrating the great potential for non-contact and non-invasive control. However, research related to the precise control of microscale actuators is still in the early stages, and in-depth research is needed for the efficient control and diversification of a range of applications. In the future, the combination of the fs laser-based fabrication technique for the precise fabrication of microscale actuators/robots and their manipulation can be established as a next-generation processing method by presenting the possibility of applications to various areas.

scholarly journals Two-photon polymerization nanolithography technology for fabrication of stimulus-responsive micro/nano-structures for biomedical applications

2020 ◽  
Vol 9 (1) ◽  
pp. 1118-1136
Author(s):  
Zhenjia Huang ◽  
Gary Chi-Pong Tsui ◽  
Yu Deng ◽  
Chak-Yin Tang
Keyword(s):  
Biomedical Applications ◽  
Three Dimensional ◽  
Water Soluble ◽  
Fabrication Technology ◽  
Nano Fabrication ◽  
Two Photon ◽  
Nano Structures ◽  
Wide Range ◽  
Stimulus Responsive ◽  
Two Photon Polymerization

AbstractMicro/nano-fabrication technology via two-photon polymerization (TPP) nanolithography is a powerful and useful manufacturing tool that is capable of generating two dimensional (2D) to three dimensional (3D) arbitrary micro/nano-structures of various materials with a high spatial resolution. This technology has received tremendous interest in cell and tissue engineering and medical microdevices because of its remarkable fabrication capability for sophisticated structures from macro- to nano-scale, which are difficult to be achieved by traditional methods with limited microarchitecture controllability. To fabricate precisely designed 3D micro/nano-structures for biomedical applications via TPP nanolithography, the use of photoinitiators (PIs) and photoresists needs to be considered comprehensively and systematically. In this review, widely used commercially available PIs are first discussed, followed by elucidating synthesis strategies of water-soluble initiators for biomedical applications. In addition to the conventional photoresists, the distinctive properties of customized stimulus-responsive photoresists are discussed. Finally, current limitations and challenges in the material and fabrication aspects and an outlook for future prospects of TPP for biomedical applications based on different biocompatible photosensitive composites are discussed comprehensively. In all, this review provides a basic understanding of TPP technology and important roles of PIs and photoresists for fabricating high-precision stimulus-responsive micro/nano-structures for a wide range of biomedical applications.

Fabrication of Three-Dimensional Micro-Structures with Two-Photon Absorption by Femtosecond Laser

2006 ◽  
Vol 532-533 ◽  
pp. 568-571
Author(s):  
Ming Zhou ◽  
Hai Feng Yang ◽  
Li Peng Liu ◽  
Lan Cai
Keyword(s):  
Photonic Crystal ◽  
Femtosecond Laser ◽  
Detection System ◽  
Three Dimensional ◽  
Photon Absorption ◽  
Micro Fabrication ◽  
Two Photon Absorption ◽  
Two Photon ◽  
Photo Polymerization ◽  
System A

The photo-polymerization induced by Two-Photon Absorption (TPA) is tightly confined in the focus because the efficiency of TPA is proportional to the square of intensity. Three-dimensional (3D) micro-fabrication can be achieved by controlling the movement of the focus. Based on this theory, a system for 3D-micro-fabrication with femtosecond laser is proposed. The system consists of a laser system, a microscope system, a real-time detection system and a 3D-movement system, etc. The precision of micro-machining reaches a level down to 700nm linewidth. The line width was inversely proportional to the fabrication speed, but proportional to laser power and NA. The experiment results were simulated, beam waist of 0.413μm and TPA cross section of 2×10-54cm4s was obtained. While we tried to optimize parameters, we also did some research about its applications. With TPA photo-polymerization by means of our experimental system, 3D photonic crystal of wood-pile structure twelve layers and photonic crystal fiber are manufactured. These results proved that the micro-fabrication system of TPA can not only obtain the resolution down to sub-micron level, but also realize real 3D micro-fabrication.

Acoustic Field-Assisted Two-Photon Polymerization Process

2021 ◽  
Vol 143 (10) ◽  
Author(s):  
Ketki M. Lichade ◽  
Yayue Pan
Keyword(s):  
Polymer Composite ◽  
Acoustic Field ◽  
Laser Power ◽  
Liquid Droplet ◽  
Three Dimensional ◽  
Polymerization Process ◽  
Photothermal Effect ◽  
Full Potential ◽  
Two Photon ◽  
Two Photon Polymerization

Abstract This study successfully integrates acoustic patterning with the Two-Photon Polymerization (TPP) process for printing nanoparticle–polymer composite microstructures with spatially varied nanoparticle compositions. Currently, the TPP process is gaining increasing attention within the engineering community for the direct manufacturing of complex three-dimensional (3D) microstructures. Yet the full potential of TPP manufactured microstructures is limited by the materials used. This study aims to create and demonstrate a novel acoustic field-assisted TPP (A-TPP) process, which can instantaneously pattern and assemble nanoparticles in a liquid droplet, and fabricate anisotropic nanoparticle–polymer composites with spatially controlled particle–polymer material compositions. It was found that the biggest challenge in integrating acoustic particle patterning with the TPP process is that nanoparticles move upon laser irradiation due to the photothermal effect, and hence, the acoustic assembly is distorted during the photopolymerization process. To cure acoustic assembly of nanoparticles in the resin through TPP with the desired nanoparticle patterns, the laser power needs to be carefully tuned so that it is adequate for curing while low enough to prevent the photothermal effect. To address this challenge, this study investigated the threshold laser power for polymerization of TPP resin (Pthr) and photothermal instability of the nanoparticle (Pthp). Patterned nanoparticle–polymer composite microstructures were fabricated using the novel A-TPP process. Experimental results validated the feasibility of the developed acoustic field-assisted TPP process on printing anisotropic composites with spatially controlled material compositions.

scholarly journals Microvascular Sprouting, Extension, and Creation of New Capillary Connections with Adaptation of the Neighboring Astrocytes in Adult Mouse Cortex under Chronic Hypoxia

2013 ◽  
Vol 34 (2) ◽  
pp. 325-331 ◽  
Author(s):  
Kazuto Masamoto ◽  
Hiroyuki Takuwa ◽  
Chie Seki ◽  
Junko Taniguchi ◽  
Yoshiaki Itoh ◽  
...  
Keyword(s):  
Adult Mouse ◽  
Fluorescent Protein ◽  
Vascular Endothelial Cells ◽  
Three Dimensional ◽  
Vascular Endothelial ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Mouse Cortex ◽  
Hypoxia Adaptation ◽  
Green Fluorescent

The present study aimed to determine the spatiotemporal dynamics of microvascular and astrocytic adaptation during hypoxia-induced cerebral angiogenesis. Adult C57BL/6J and Tie2-green fluorescent protein (GFP) mice with vascular endothelial cells expressing GFP were exposed to normobaric hypoxia for 3 weeks, whereas the three-dimensional microvessels and astrocytes were imaged repeatedly using two-photon microscopy. After 7 to14 days of hypoxia, a vessel sprout appeared from the capillaries with a bump-like head shape (mean diameter 14  μm), and stagnant blood cells were seen inside the sprout. However, no detectable changes in the astrocyte morphology were observed for this early phase of the hypoxia adaptation. More than 50% of the sprouts emerged from capillaries 60  μm away from the center penetrating arteries, which indicates that the capillary distant from the penetrating arteries is a favored site for sprouting. After 14 to 21 days of hypoxia, the sprouting vessels created a new connection with an existing capillary. In this phase, the shape of the new vessel and its blood flow were normalized, and the outside of the vessels were wrapped with numerous processes from the neighboring astrocytes. The findings indicate that hypoxia-induced cerebral angiogenesis provokes the adaptation of neighboring astrocytes, which may stabilize the blood–brain barrier in immature vessels.

scholarly journals Rapid Fabrication of Continuous Surface Fresnel Microlens Array by Femtosecond Laser Focal Field Engineering

Micromachines ◽  
2020 ◽  
Vol 11 (2) ◽  
pp. 112 ◽  
Author(s):  
Linyu Yan ◽  
Dong Yang ◽  
Qihuang Gong ◽  
Yan Li
Keyword(s):  
Femtosecond Laser ◽  
Three Dimensional ◽  
Fresnel Lens ◽  
Two Dimensional ◽  
Direct Writing ◽  
Lens Array ◽  
Two Photon ◽  
Rapid Fabrication ◽  
Continuous Surface ◽  
Two Photon Polymerization

Femtosecond laser direct writing through two-photon polymerization has been widely used in precision fabrication of three-dimensional microstructures but is usually time consuming. In this article, we report the rapid fabrication of continuous surface Fresnel lens array through femtosecond laser three-dimensional focal field engineering. Each Fresnel lens is formed by continuous two-photon polymerization of the two-dimensional slices of the whole structure with one-dimensional scan of the corresponding two-dimensional engineered intensity distribution. Moreover, we anneal the lens array to improve its focusing and imaging performance.

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