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Two photon microscopy

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https://www.readbyqxmd.com/read/28736661/brain-tumor-delineation-enhanced-by-moxifloxacin-based-two-photon-cars-combined-microscopy
#1
Viet-Hoan Le, Su Woong Yoo, Yeoreum Yoon, Taejun Wang, Bumju Kim, Seunghun Lee, Kyung-Hwa Lee, Ki Hean Kim, Euiheon Chung
Delineating brain tumor margin is critical for maximizing tumor removal while sparing adjacent normal tissue for better clinical outcome. We describe the use of moxifloxacin-based two-photon (TP)/coherent anti-Stokes Raman scattering (CARS) combined microscopy for differentiating normal mouse brain tissue from metastatic brain tumor tissue based on histoarchitectural and biochemical differences. Moxifloxacin, an FDA-approved compound, was used to label cells in the brain, and moxifloxacin-based two-photon microscopy (TPM) revealed tumor lesions with significantly high cellular density and invading edges in a metastatic brain tumor model...
April 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/28734477/polarity-sensitive-probes-for-superresolution-stimulated-emission-depletion-microscopy
#2
Erdinc Sezgin, Falk Schneider, Victoria Zilles, Iztok Urbančič, Esther Garcia, Dominic Waithe, Andrey S Klymchenko, Christian Eggeling
The lateral organization of molecules in the cellular plasma membrane plays an important role in cellular signaling. A critical parameter for membrane molecular organization is how the membrane lipids are packed. Polarity-sensitive dyes are powerful tools to characterize such lipid membrane order, employing, for example, confocal and two-photon microscopy. The investigation of potential nanodomains, however, requires the use of superresolution microscopy. Here, we test the performance of the polarity-sensitive membrane dyes Di-4-ANEPPDHQ, Di-4-AN(F)EPPTEA, and NR12S in superresolution stimulated emission depletion microscopy...
July 19, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/28726383/marangoni-convection-assisted-single-molecule-detection-with-nanojet-surface-enhanced-raman-spectroscopy
#3
Te-Wei Chang, Xinhao Wang, Amirreza Mahigir, Georgios Veronis, Gang Logan Liu, Manas Ranjan Gartia
Many single-molecule (SM) label-free techniques such as scanning probe microscopies (SPM) and magnetic force spectroscopies (MFS) provide high resolution surface topography information, but lack chemical information. Typical surface enhanced Raman spectroscopy (SERS) systems provide chemical information on the analytes, but lack spatial resolution. In addition, a challenge in SERS sensors is to bring analytes into the so-called "hot spots" (locations where the enhancement of electromagnetic field amplitude is larger than 10(3))...
July 20, 2017: ACS Sensors
https://www.readbyqxmd.com/read/28717572/multiphoton-microscopy-for-label-free-identification-of-intramural-metastasis-in-human-esophageal-squamous-cell-carcinoma
#4
Jian Xu, Deyong Kang, Yaping Zeng, Shuangmu Zhuo, Xiaoqin Zhu, Liwei Jiang, Jianxin Chen, Jiangbo Lin
For complete removal of cancerous tissue in esophageal squamous cell carcinoma (ESCC), intramural metastasis (IM) should be identified preoperatively or intraoperatively. Here, multiphoton microscopy (MPM) was introduced for label-free identification of IM in the esophageal wall, by a combination of two-photon excited fluorescence (TPEF), second harmonic generation (SHG) imaging, and spectral analysis. Three-dimensional (3D) imaging of the IM region was also performed. Quantitative parameters, including 3D fiber orientation, were measured by 3D-weighted orientation vector summation...
July 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/28717558/single-pulse-two-photon-fluorescence-lifetime-imaging-sp-flim-with-mhz-pixel-rate
#5
Matthias Eibl, Sebastian Karpf, Daniel Weng, Hubertus Hakert, Tom Pfeiffer, Jan Philip Kolb, Robert Huber
Two-photon-excited fluorescence lifetime imaging microscopy (FLIM) is a chemically specific 3-D sensing modality providing valuable information about the microstructure, composition and function of a sample. However, a more widespread application of this technique is hindered by the need for a sophisticated ultra-short pulse laser source and by speed limitations of current FLIM detection systems. To overcome these limitations, we combined a robust sub-nanosecond fiber laser as the excitation source with high analog bandwidth detection...
July 1, 2017: Biomedical Optics Express
https://www.readbyqxmd.com/read/28716943/visualizing-context-dependent-calcium-signaling-in-encephalitogenic-t-cells-in-vivo-by-two-photon-microscopy
#6
Nikolaos I Kyratsous, Isabel J Bauer, Guokun Zhang, Marija Pesic, Ingo Bartholomäus, Marsilius Mues, Ping Fang, Miriam Wörner, Stephanie Everts, Joachim W Ellwart, Joanna M Watt, Barry V L Potter, Reinhard Hohlfeld, Hartmut Wekerle, Naoto Kawakami
In experimental autoimmune encephalitis (EAE), autoimmune T cells are activated in the periphery before they home to the CNS. On their way, the T cells pass through a series of different cellular milieus where they receive signals that instruct them to invade their target tissues. These signals involve interaction with the surrounding stroma cells, in the presence or absence of autoantigens. To portray the serial signaling events, we studied a T-cell-mediated model of EAE combining in vivo two-photon microscopy with two different activation reporters, the FRET-based calcium biosensor Twitch1 and fluorescent NFAT...
July 17, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28710672/automated-3-d-detection-of-dendritic-spines-from-in-vivo-two-photon-image-stacks
#7
P K Singh, P Hernandez-Herrera, D Labate, M Papadakis
Despite the significant advances in the development of automated image analysis algorithms for the detection and extraction of neuronal structures, current software tools still have numerous limitations when it comes to the detection and analysis of dendritic spines. The problem is especially challenging in in vivo imaging, where the difficulty of extracting morphometric properties of spines is compounded by lower image resolution and contrast levels native to two-photon laser microscopy. To address this challenge, we introduce a new computational framework for the automated detection and quantitative analysis of dendritic spines in vivo multi-photon imaging...
July 14, 2017: Neuroinformatics
https://www.readbyqxmd.com/read/28709993/intracellular-calcium-release-through-ip3r-or-ryr-contributes-to-secondary-axonal-degeneration
#8
Ben C Orem, Nicolas Pelisch, Joshua Williams, Jacqueline M Nally, David P Stirling
Severed CNS axons often retract or dieback away from the injury site and fail to regenerate. The precise mechanisms underlying acute axonal dieback and secondary axonal degeneration remain poorly understood. Here we investigate the role of Ca(2+) store mediated intra-axonal Ca(2+) release in acute axonal dieback and secondary axonal degeneration. To differentiate between primary (directly transected) and "bystander" axonal injury (axons spared by the initial injury but then succumb to secondary degeneration) in real-time we use our previously published highly focal laser-induced spinal cord injury (LiSCI) ex vivo model...
July 12, 2017: Neurobiology of Disease
https://www.readbyqxmd.com/read/28706994/short-range-surface-plasmonics-localized-electron-emission-dynamics-from-a-60-nm-spot-on-an-atomically-flat-single-crystalline-gold-surface
#9
Bettina Frank, Philip Kahl, Daniel Podbiel, Grisha Spektor, Meir Orenstein, Liwei Fu, Thomas Weiss, Michael Horn-von Hoegen, Timothy J Davis, Frank-J Meyer Zu Heringdorf, Harald Giessen
We experimentally and theoretically visualize the propagation of short-range surface plasmon polaritons using atomically flat single-crystalline gold platelets on silicon substrates. We study their excitation and subfemtosecond dynamics via normal-incidence two-photon photoemission electron microscopy. By milling a plasmonic disk and grating structure into a single-crystalline gold platelet, we observe nanofocusing of the short-range surface plasmon polariton. Localized two-photon ultrafast electron emission from a spot with a smallest dimension of 60 nm is observed...
July 2017: Science Advances
https://www.readbyqxmd.com/read/28700127/enhanced-volumetric-imaging-in-two-photon-microscopy-via-acoustic-lens-beam-shaping
#10
Simonluca Piazza, Paolo Bianchini, Colin Sheppard, Alberto Diaspro, Martí Duocastella
Three-dimensional imaging at high-spatiotemporal resolutions and over large penetration depths is key for unmasking the dynamics and structural organization of complex biological systems. However, the need to axially shift the focus, with consequent limitations in imaging speed, and signal degradation at large depths due to scattering effects, makes this task challenging. Here, we present a novel approach in two-photon excitation microscopy that allows fast volumetric imaging and enhanced signal-to-background (S/B) in thick tissue...
July 12, 2017: Journal of Biophotonics
https://www.readbyqxmd.com/read/28697466/a-ratiometric-two-photon-probe-for-ca-2-in-live-tissues-and-its-application-to-spinal-cord-injury-model
#11
Hyung Joong Kim, Chang Su Lim, Hyo Won Lee, Hye Sue Lee, Yun Ju Um, Hemant Kumar, Inbo Han, Hwan Myung Kim
Ratiometric imaging with a small-molecule probe is important for the in-situ quantitative analysis of chemical events. We developed a ratiometric two-photon fluorescent probe (SCa1-IREF) derived from dual dyes with different Stokes shifts. This probe has two identical windows: a Ca(2+)-sensing window and an internal reference window, with eliminated FRET interference. SCa1-IREF shows a marked change in the ratio upon response with Ca(2+), significant two-photon brightness, considerable selectivity for Ca(2+), and cell loading ability with low cytotoxicity...
July 6, 2017: Biomaterials
https://www.readbyqxmd.com/read/28694464/monocular-deprivation-induces-dendritic-spine-elimination-in-the-developing-mouse-visual-cortex
#12
Yanmei Zhou, Baoling Lai, Wen-Biao Gan
It is well established that visual deprivation has a profound impact on the responsiveness of neurons in the developing visual cortex. The effect of visual deprivation on synaptic connectivity remains unclear. Using transcranial two-photon microscopy, we examined the effect of visual deprivation and subsequent recovery on dendritic spine remodeling of layer 5 pyramidal neurons in the mouse primary visual cortex. We found that monocular deprivation (MD), but not binocular deprivation (BD), increased dendritic spine elimination over 3 days in the binocular region of 4-week-old adolescent mice...
July 10, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28686582/measurement-of-drug-target-engagement-in-live-cells-by-two-photon-fluorescence-anisotropy-imaging
#13
Claudio Vinegoni, Paolo Fumene Feruglio, Christian Brand, Sungon Lee, Antoinette E Nibbs, Shawn Stapleton, Sunil Shah, Ignacy Gryczynski, Thomas Reiner, Ralph Mazitschek, Ralph Weissleder
The ability to directly image and quantify drug-target engagement and drug distribution with subcellular resolution in live cells and whole organisms is a prerequisite to establishing accurate models of the kinetics and dynamics of drug action. Such methods would thus have far-reaching applications in drug development and molecular pharmacology. We recently presented one such technique based on fluorescence anisotropy, a spectroscopic method based on polarization light analysis and capable of measuring the binding interaction between molecules...
July 2017: Nature Protocols
https://www.readbyqxmd.com/read/28685439/brain-tissue-po2-measurement-during-normoxia-and-hypoxia-using-two-photon-phosphorescence-lifetime-microscopy
#14
Kui Xu, David A Boas, Sava Sakadžić, Joseph C LaManna
Key to the understanding of the principles of physiological and structural acclimatization to changes in the balance between energy supply (represented by substrate and oxygen delivery, and mitochondrial oxidative phosphorylation) and energy demand (initiated by neuronal activity) is to determine the controlling variables, how they are sensed and the mechanisms initiated to maintain the balance. The mammalian brain depends completely on continuous delivery of oxygen to maintain its function. We hypothesized that tissue oxygen is the primary sensed variable...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28684735/measurement-of-nanoscale-three-dimensional-diffusion-in-the-interior-of-living-cells-by-sted-fcs
#15
Luca Lanzanò, Lorenzo Scipioni, Melody Di Bona, Paolo Bianchini, Ranieri Bizzarri, Francesco Cardarelli, Alberto Diaspro, Giuseppe Vicidomini
The observation of molecular diffusion at different spatial scales, and in particular below the optical diffraction limit (<200 nm), can reveal details of the subcellular topology and its functional organization. Stimulated-emission depletion microscopy (STED) has been previously combined with fluorescence correlation spectroscopy (FCS) to investigate nanoscale diffusion (STED-FCS). However, stimulated-emission depletion fluorescence correlation spectroscopy has only been used successfully to reveal functional organization in two-dimensional space, such as the plasma membrane, while, an efficient implementation for measurements in three-dimensional space, such as the cellular interior, is still lacking...
July 6, 2017: Nature Communications
https://www.readbyqxmd.com/read/28674487/pyramidal-neurons-in-different-cortical-layers-exhibit-distinct-dynamics-and-plasticity-of-apical-dendritic-spines
#16
Michelle Tjia, Xinzhu Yu, Lavpreet S Jammu, Ju Lu, Yi Zuo
The mammalian cerebral cortex is typically organized in six layers containing multiple types of neurons, with pyramidal neurons (PNs) being the most abundant. PNs in different cortical layers have distinct morphology, physiology and functional roles in neural circuits. Therefore, their development and synaptic plasticity may also differ. Using in vivo transcranial two-photon microscopy, we followed the structural dynamics of dendritic spines on apical dendrites of layer (L) 2/3 and L5 PNs at different developmental stages...
2017: Frontiers in Neural Circuits
https://www.readbyqxmd.com/read/28672005/fatty-acid-transport-protein-1-regulates-retinoid-metabolism-and-photoreceptor-development-in-mouse-retina
#17
Aurélie Cubizolle, Laurent Guillou, Bertrand Mollereau, Christian P Hamel, Philippe Brabet
In retinal pigment epithelium (RPE), RPE65 catalyzes the isomerization of all-trans-retinyl fatty acid esters to 11-cis-retinol in the visual cycle and controls the rhodopsin regeneration rate. However, the mechanisms by which these processes are regulated are still unclear. Fatty Acid Transport Protein 1 (FATP1) is involved in fatty acid uptake and lipid metabolism in a variety of cell types. FATP1 co-localizes with RPE65 in RPE and inhibits its isomerase activity in vitro. Here, we further investigated the role of FATP1 in the visual cycle using transgenic mice that overexpress human FATP1 specifically in the RPE (hFATP1TG mice)...
2017: PloS One
https://www.readbyqxmd.com/read/28669077/lung-as-a-niche-for-hematopoietic-progenitors
#18
Isabella Borges, Isadora Sena, Patrick Azevedo, Julia Andreotti, Viviani Almeida, Ana Paiva, Gabryella Santos, Daniel Guerra, Pedro Prazeres, Luiza Lousado Mesquita, Luanny Souto de Barros Silva, Caroline Leonel, Akiva Mintz, Alexander Birbrair
Platelets are released from megakaryocytes. The bone marrow has been proposed to be the major site where this process occurs. Lefrançais et al. (2017) using state-of-the-art techniques including two-photon microscopy, in vivo lineage-tracing technologies, and sophisticated lung transplants reveal that the lung is also a primary site for platelet biogenesis. Strikingly, lung megakaryocytes can completely reconstitute platelet counts in the blood in mice with thrombocytopenia. This study also shows that hematopoietic progenitors, with capacity to repopulate the bone marrow after irradiation, are present in the lungs...
July 1, 2017: Stem Cell Reviews
https://www.readbyqxmd.com/read/28668578/computer-assisted-three-dimensional-tracking-of-sensory-innervation-in-the-murine-bladder-mucosa-with-two-photon-microscopy
#19
Anna Schueth, Bart Spronck, Marc A M J van Zandvoort, Gommert A van Koeveringe
A strong association between functional bladder disorders and bladder sensation is well-known, with a relationship between malfunctioning detrusor muscle and abnormal sensation arising from the sub-urothelium and the lamina propria (LP), has been suggested. However, the exact underlying pathophysiology of these bladder disorders is not completely understood. Therefore, it is important to gain knowledge on sensory innervation of the urinary bladder in order to understand the neural network function in healthy and diseased bladder...
June 28, 2017: Journal of Chemical Neuroanatomy
https://www.readbyqxmd.com/read/28668541/non-linear-optical-microscopy-as-a-novel-quantitative-and-label-free-imaging-modality-to-improve-the-assessment-of-tissue-engineered-cartilage-tec
#20
A Islam, E I Romijn, M B Lilledahl, I Martinez-Zubiaurre
OBJECTIVE: Current systems to evaluate outcomes from tissue-engineered cartilage (TEC) are sub-optimal. The main purpose of our study was to demonstrate the use of second harmonic generation (SHG) microscopy as a novel quantitative approach to assess collagen deposition in laboratory made cartilage constructs. METHODS: Scaffold-free cartilage constructs were obtained by condensation of in vitro expanded Hoffa's fat pad derived stromal cells (HFPSCs), incubated in the presence or absence of chondrogenic growth factors (GF) during a period of 21 d...
June 29, 2017: Osteoarthritis and Cartilage
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