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1.
J Biophotonics ; 17(4): e202300422, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38211977

RESUMEN

Multiphoton microscopy (MPM) enables deep brain imaging. Three optical windows: NIR-I, NIR-II, and NIR-III are widely used. Recently, NIR-IV (the 2200 nm window) has been demonstrated to be the last and longest window for deep tissue MPM. However, so far MPM covers only two optical windows labeled by single fluorescent probe, one for emission and one for excitation. Here we demonstrate in vivo deep brain MPM covering three optical windows, with emission at NIR-I, NIR-II, and excitation at NIR-IV, labeled by ICG. The innovations include: (1) characterizing both 3-photon excitation and emission properties of ICG emitting at both NIR-I and NIR-II, in water, plasma, and circulating blood; (2) a home-built multiphoton microscope with simultaneous dual channel detection, with which we demonstrate deep brain MPM 950 µm (NIR-I) and 850 µm (NIR-II) into the mouse brain in vivo, verifying that multi-optical window MPM is promising for deep brain imaging.


Asunto(s)
Encéfalo , Microscopía de Fluorescencia por Excitación Multifotónica , Ratones , Animales , Microscopía de Fluorescencia por Excitación Multifotónica/métodos , Encéfalo/diagnóstico por imagen , Colorantes Fluorescentes , Imagen Óptica/métodos , Espectroscopía Infrarroja Corta/métodos
2.
Huan Jing Ke Xue ; 44(4): 1998-2008, 2023 Apr 08.
Artículo en Chino | MEDLINE | ID: mdl-37040950

RESUMEN

Focused on the key areas of energy, buildings, industry, and transportation, with 2020 as the base year and 2035 as the target year, we respectively designed the baseline scenario, policy scenario, and enhanced scenario, calculated the emission reduction potential of air pollutants and CO2 of Beijing, and constructed an assessment method of co-control effect gradation index to evaluate the co-control effect of air pollutants and CO2 in the policy scenario and enhanced scenario. The results showed that in the policy scenario and enhanced scenario, the reduction rates of air pollutants emissions will reach 11%-75% and 12%-94%, respectively, and reduction rates of CO2 emissions will reach 41% and 52%, respectively, compared with those from the baseline scenario. Optimizing vehicle structure had the largest contribution to the emission reduction of NOx, VOCs, and CO2, and the emission reduction rates will reach 74%, 80%, and 31% in the policy scenario and 68%, 74%, and 22% in the enhanced scenario, respectively. Replacing coal-fired with clean energy in rural areas had the largest contribution to the emission reduction of SO2; the emission reduction rates will reach 47% and 35% in the policy scenario and enhanced scenario, respectively. Improving the green level of new buildings had the largest contribution to the emission reduction of PM10; the emission reduction rates will reach 79% and 74% in the policy scenario and enhanced scenario, respectively. Optimizing travel structure and promoting green development of digital infrastructure had the best co-control effect. The co-control effect of replacing coal-fired with clean energy in rural areas, optimizing vehicle structure, and promoting green upgrading of the manufacturing industry will be improved to a better status in the enhanced scenario. More attention should be paid to improving the proportion of green trips, implementing the promotion of new energy vehicles, and the green transportation of goods to reduce emissions in the field of transportation. At the same time, with the continuous improvement in electrification level in the end energy consumption structure, the proportion of green electricity should be increased by expanding local renewable energy power production and increasing external green electricity transmission capacity, to enhance the co-control effect of pollution and carbon reduction.

3.
ACS Nano ; 17(4): 3686-3695, 2023 02 28.
Artículo en Inglés | MEDLINE | ID: mdl-36799427

RESUMEN

Multiphoton microscopy (MPM) is an enabling technology for visualizing deep-brain structures at high spatial resolution in vivo. Within the low tissue absorption window, shifting to longer excitation wavelengths reduces tissue scattering and boosts penetration depth. Recently, the 2200 nm excitation window has emerged as the last and longest window suitable for deep-brain MPM. However, multiphoton fluorescence imaging at this window has not been demonstrated, due to the lack of characterization of multiphoton properties of fluorescent labels. Here we demonstrate technologies for measuring both the multiphoton excitation and emission properties of fluorescent labels at the 2200 nm window, using (1) 3-photon (ησ3) and 4-photon action cross sections (ησ4) and (2) 3-photon and 4-photon emission spectra both ex vivo and in vivo of quantum dots. Our results show that quantum dots have exceptionally large ησ3 and ησ4 for efficient generation of multiphoton fluorescence. Besides, the 3-photon and 4-photon emission spectra of quantum dots are essentially identical to those of one-photon emission, which change negligibly subject to the local environment of circulating blood. Based on these characterization results, we further demonstrate deep-brain vasculature imaging in vivo. Due to the superb multiphoton properties of quantum dots, 3-photon and 4-photon fluorescence imaging reaches a maximum brain imaging depth of 1060 and 940 µm below the surface of a mouse brain, respectively, which enables the imaging of subcortical structures. We thus fill the last gap in multiphoton fluorescence imaging in terms of wavelength selection.


Asunto(s)
Puntos Cuánticos , Animales , Ratones , Puntos Cuánticos/química , Encéfalo/diagnóstico por imagen , Encéfalo/irrigación sanguínea , Microscopía de Fluorescencia por Excitación Multifotónica/métodos , Colorantes Fluorescentes/química , Imagen Óptica
4.
J Biophotonics ; 16(5): e202200365, 2023 05.
Artículo en Inglés | MEDLINE | ID: mdl-36633161

RESUMEN

The brain arteriolar wall is a multilayered structure, whose integrity is of key significance to the brain function. However, resolving these different layers in anmial models in vivo is hampered by the lack of either labeling or imaging technology. Here, we demonstrate that three-photon microscopy (3PM) is an ideal solution. In mouse brain in vivo, excited at the 1700-nm window, label-free third-harmonic generation imaging and three-photon fluorescence (3PF) imaging with Alexa 633 labeling colocalize and resolve the internal elastic lamina. Furthermore, Alexa Fluor 594-conjugated Wheat Germ Agglutinin (WGA-594) shows time-dependent labeling behavior. As time lapses, WGA-594 first labels endothelium, and then vascular smooth muscle cells, which are readily captured and resolved with 3PF imaging. Our results show that 3PM, in combination with proper labeling, is a promising technology for investigating the structures of brain arteriolar wall in vivo.


Asunto(s)
Encéfalo , Microscopía de Fluorescencia por Excitación Multifotónica , Ratones , Animales , Encéfalo/diagnóstico por imagen , Microscopía de Fluorescencia por Excitación Multifotónica/métodos , Endotelio
5.
Biomed Opt Express ; 12(10): 6474-6484, 2021 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-34745750

RESUMEN

Visualization of lymphatic vessels is key to the understanding of their structure, function, and dynamics. Multiphoton microscopy (MPM) is a potential technology for imaging lymphatic vessels, but tissue scattering prevents its deep penetration in skin. Here we demonstrate deep-skin MPM of the lymphatic vessels in mouse hindlimb in vivo, excited at the 1700 nm window. Our results show that with contrast provided by indocyanine green (ICG), 2-photon fluorescence (2PF) imaging enables noninvasive imaging of lymphatic vessels 300 µm below the skin surface, visualizing both its structure and contraction dynamics. Simultaneously acquired second-harmonic generation (SHG) and third-harmonic generation (THG) images visualize the local environment in which the lymphatic vessels reside. After removing the surface skin layer, 2PF and THG imaging visualize finer structures of the lymphatic vessels: most notably, the label-free THG imaging visualizes lymphatic valves and their open-and-close dynamics in real time. MPM excited at the 1700-nm window thus provides a promising technology for the study of lymphatic vessels.

6.
Rev Sci Instrum ; 92(5): 055102, 2021 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-34243303

RESUMEN

Rock burst early warning technology is currently applied mainly in microseismic monitoring. Rock burst signals indicate the micro-fracture phenomena of a rock and can transmit earthquake waves through the rock before they are finally received by a detector. A characteristic decomposition of rock micro-fracture signals was conducted by the singular value Empirical Mode Decomposition (EMD) algorithm to effectively decompose the characteristic signals of a rock micro-fracture from mixed microseismic signals, with a low signal to noise ratio to ensure prediction precision. When comparing the proposed method with wavelet decomposition and EMD, it was found that the local characteristics of the signals were retained effectively. The proposed algorithm was verified by applying it in a laboratory simulation and to the decomposition of microseismic signals from a hydro-power station. It was concluded that the improved algorithm had a better decomposition precision than wavelet decomposition and EMD decomposition and could effectively separate the characteristic signals of micro-earthquakes. This could provide a significant basis for the identification of the abnormal microseismic signals of rock micro-fractures as well as a pre-warning of rock fractures. It is therefore of practical significance to study rock fracture early warning technology.

7.
J Biophotonics ; 14(10): e202100076, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34160142

RESUMEN

Multiphoton microscopy (MPM) excited at the 1700-nm window has enabled deep-tissue penetration in biological tissue, especially brain. MPM of skin may also benefit from this deep-penetration capability. Skin is a layered structure with varying refractive index (from 1.34 to 1.5). Consequently, proper immersion medium should be selected when imaging with high numerical aperture objective lens. To provide guidelines for immersion medium selection for skin MPM, here we demonstrate comparative experimental investigation of deep-skin MPM excited at 1600 nm in vivo, using both silicone oil and deuterium dioxide (D2 O) immersion. We specifically characterize imaging depths, signal levels and spatial resolution. Our results show that both immersion media give similar performance in imaging depth and spatial resolution, while signal levels are slightly better with silicone oil immersion. We also demonstrate that local injection of fluorescent beads into the skin is a viable technique for spatial resolution characterization in vivo.


Asunto(s)
Microscopía de Fluorescencia por Excitación Multifotónica , Aceites de Silicona , Deuterio , Inmersión , Piel/diagnóstico por imagen
8.
J Biophotonics ; 14(3): e202000349, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33179837

RESUMEN

3-photon microscopy (3PM) excited at the 1700 nm window enables deep-tissue imaging in vivo, especially in brain. PC rod soliton source has previously been exclusively used as the excitation source, which is rather costly and difficult to align. Here we demonstrate a novel nonlinear optical technique to build femtosecond laser source at the 1700 nm window, based on self-phase modulation (SPM) in a short span of large-mode-area fiber. The spectral broadening experienced by the pump pulse leads to the generation of a red-shifted sidelobe at 1603 nm. After spectral filtering, this sidelobe corresponds to 170-fs, 167-nJ pulses at 1603 nm. Using this SPM source, we further demonstrate deep-brain 3 PM to a depth of 1500 µm below the mouse brain surface in vivo. Our SPM femtosecond laser source thus provides a cost effective and easy-to-align alternative excitation source to the PC rod soliton source.


Asunto(s)
Microscopía , Fotones , Animales , Encéfalo/diagnóstico por imagen , Rayos Láser , Ratones
9.
Opt Lett ; 45(18): 5271-5274, 2020 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-32932509

RESUMEN

Microglia act as the first and main form of active immune defense in brain. However, in animal models, research on these cells is limited to the superficial layer of the brain, due to the lack of a deep-imaging technique. Here we break this depth limit using three-photon fluorescence (3PF) microscopy excited at the 1700-nm window. Three-photon action cross-section (ησ3) measurement lays the basis for dye selection and the resultant maximization of 3PF generation. 3PF imaging suppresses the surface background, leading to a much improved signal-to-background ratio compared to the commonly used two-photon microscopy (2PM). We can image microglia 1124 µm below the brain surface in vivo, 3.7 times deeper than previous results using 2PM for microglia imaging. This technique enables us to visualize microglia in the white matter layer in vivo for the first time.


Asunto(s)
Encéfalo/citología , Encéfalo/diagnóstico por imagen , Microglía/citología , Microscopía de Fluorescencia por Excitación Multifotónica , Animales , Ratones , Relación Señal-Ruido
10.
J Biophotonics ; 13(12): e202000321, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-32969170

RESUMEN

Myelin is a key component of the peripheral nervous system, whose structure anomaly in the digital skin is implicated in neuropathy. Here we demonstrate an in vivo labeling and imaging technique, capable of visualizing myelin sheaths deep in the mouse digital skin. Through material characterization, we verify that 3-photon fluorescence (3PF) can be generated from a commonly used dye- FluoroMyelin Red for labeling myelin, excited at the 1700-nm window. Topical injection of FluoroMyelin Red in the mouse digit leads to bright labeling of myelin sheaths. Harnessing the deep-penetration capability of 3-photon microscopy excited at the 1700-nm window, we demonstrate that 3PF imaging of FluoroMyelin Red-labeled myelin sheaths in the mouse digit in vivo can be achieved to a depth 340 µm below the skin surface, revealing both branching bundle of and individual myelin sheaths.


Asunto(s)
Microscopía , Vaina de Mielina , Animales , Ratones , Fotones
11.
Biomed Opt Express ; 11(5): 2738-2744, 2020 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-32499956

RESUMEN

Measurement of the hemodynamic physical parameter blood flow speed in the brain in vivo is key to understanding brain physiology and pathology. 2-photon fluorescence microscopy with single blood vessel resolution is typically used, which necessitates injection of toxic fluorescent dyes. Here we demonstrate a label-free nonlinear optical technique, third-harmonic generation microscopy excited at the 1700-nm window, that is promising for such measurement. Using a simple femtosecond laser system based on soliton self-frequency shift, we can measure blood flow speed through the whole cortical grey matter, even down to the white matter layer. Together with 3-photon fluorescence microscopy, we further demonstrate that the blood vessel walls generate strong THG signals, and that plasma and circulating blood cells are mutually exclusive in space. This technique can be readily applied to brain research.

12.
Molecules ; 24(23)2019 Nov 25.
Artículo en Inglés | MEDLINE | ID: mdl-31775363

RESUMEN

In previous work, we applied the rotation-limiting strategy and introduced a substituent at the 3-position of the pyrazolo [3,4-d]pyrimidin-4-amine as the affinity element to interact with the deeper hydrophobic pocket, discovered a series of novel quinazolinones as potent PI3Kδ inhibitors. Among them, the indole derivative 3 is one of the most selective PI3Kδ inhibitors and the 3,4-dimethoxyphenyl derivative 4 is a potent and selective dual PI3Kδ/γ inhibitor. In this study, we replaced the carbonyl group in the quinazolinone core with a sulfonyl group, designed a series of novel 2H-benzo[e][1,2,4]thiadiazine 1,1-dioxide derivatives as PI3Kδ inhibitors. After the reduction of nitro group in N-(2,6-dimethylphenyl)-2-nitrobenzenesulfonamide 5 and N-(2,6-dimethylphenyl)-2-nitro-5-fluorobenzenesulfonamide 6, the resulting 2-aminobenzenesulfonamides were reacted with trimethyl orthoacetate to give the 3-methyl-2H-benzo[e][1,2,4]thiadiazine 1,1-dioxide derivatives. After bromination of the 3-methyl group, the nucleophilic substitution with the 3-iodo-1H-pyrazolo[3,4-d]pyrimidin-4-amine provided the respective iodide derivatives, which were further reacted with a series of arylboronic acids via Suzuki coupling to furnish the 2H-benzo[e][1,2,4]thiadiazine 1,1-dioxide derivatives 15a-J and 16a-d. In agreement with the quinazolinone derivatives, the introduction of a 5-indolyl or 3,4-dimethoxyphenyl at the affinity pocket generated the most potent analogues 15a and 15b with the IC50 values of 217 to 266 nM, respectively. In comparison with the quinazolinone lead compounds 3 and 4, these 2H-benzo[e][1,2,4]thiadiazine 1,1-dioxide derivatives exhibited much decreased PI3Kδ inhibitory potency, but maintained the high selectivity over other PI3K isoforms. Unlike the quinazolinone lead compound 4 that was a dual PI3Kδ/γ inhibitor, the benzthiadiazine 1,1-dioxide 15b with the same 3,4-dimethoxyphenyl moiety was more than 21-fold selective over PI3Kγ. Moreover, the introducing of a fluorine atom at the 7-position of the 2H-benzo[e][1,2,4]thiadiazine 1,1-dioxide core, in general, was not favored for the PI3Kδ inhibitory activity. In agreement with their high PI3Kδ selectivity, 15a and 15b significantly inhibited the SU-DHL-6 cell proliferation.


Asunto(s)
Fosfatidilinositol 3-Quinasa Clase I/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/síntesis química , Quinazolinonas/química , Tiadiazinas/síntesis química , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Fosfatidilinositol 3-Quinasa Clase I/química , Diseño de Fármacos , Humanos , Simulación del Acoplamiento Molecular , Neoplasias/tratamiento farmacológico , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/farmacología , Quinazolinonas/síntesis química , Quinazolinonas/farmacología , Relación Estructura-Actividad , Tiadiazinas/química , Tiadiazinas/farmacología
13.
Opt Lett ; 44(17): 4432-4435, 2019 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-31465421

RESUMEN

Here we demonstrate deep-brain 2-photon fluorescence microscopy in mouse in vivo, excited at the 1700 nm window. Through signal versus power measurement, we show that indocyanine green (ICG) is a promising 2-photon fluorescent dye excitable at the 1700 nm window. In order to excite ICG circulating in the vasculature in the deep brain, we employ a circular-polarization soliton self-frequency shift technique to generate energetic femtosecond pulses at 1617 nm. Combining the labeling and laser technologies, we achieve a record 2-photon fluorescence brain vasculature imaging depth of 2000 µm in vivo. Both the effective attenuation length measurement and signal-to-background ratio measurement indicate that we have reached the theoretical depth limit in 2-photon fluorescence microscopy.

14.
Anal Chem ; 91(15): 9371-9375, 2019 08 06.
Artículo en Inglés | MEDLINE | ID: mdl-31339298

RESUMEN

Optical access of a mouse brain using microscopes is the key to study brain structures and functions in vivo. However, the opaque skull of a mouse has to be either opened or thinned in an invasive way to attain an adequate imaging depth in the brain. Mild skull optical clearing is highly desired, but its chemical mechanism is far from being understood. Here, we unraveled the molecular process underlying optical clearing of the mouse skull by label-free hyperspectral stimulated Raman scattering (SRS) microscopy, thereby discovering the optimal clearing strategy to turn a turbid skull into a transparent skull window. Furthermore, we demonstrated in vivo three-photon imaging of vascular structures as deep as 850 µm in the cortex of the mouse brain. Coherent Raman based microspectroscopy holds great promise to advance skull and tissue clearing methods in the future.


Asunto(s)
Neuroimagen/métodos , Espectrometría Raman/métodos , Animales , Encéfalo/irrigación sanguínea , Encéfalo/citología , Encéfalo/diagnóstico por imagen , Corteza Cerebral/irrigación sanguínea , Corteza Cerebral/diagnóstico por imagen , Ratones , Cráneo
15.
Nano Lett ; 19(8): 5260-5265, 2019 08 14.
Artículo en Inglés | MEDLINE | ID: mdl-31268725

RESUMEN

Visualizing deep-brain vasculature and hemodynamics is key to understanding brain physiology and pathology. Among the various adopted imaging modalities, multiphoton microscopy (MPM) is well-known for its deep-brain structural and hemodynamic imaging capability. However, the largest imaging depth in MPM is limited by signal depletion in the deep brain. Here we demonstrate that quantum dots are an enabling material for significantly deeper structural and hemodynamic MPM in mouse brain in vivo. We characterized both three-photon excitation and emission parameters for quantum dots: the measured three-photon cross sections of quantum dots are 4-5 orders of magnitude larger than those of conventional fluorescent dyes excited at the 1700 nm window, while the three-photon emission spectrum measured in the circulating blood in vivo shows a slight red shift and broadening compared with ex vivo measurement. On the basis of these measured results, we further demonstrate both structural and hemodynamic three-photon microscopy in the mouse brain in vivo labeled by quantum dots, at record depths among all MPM modalities at all demonstrated excitation wavelengths.


Asunto(s)
Encéfalo/irrigación sanguínea , Hemodinámica , Microscopía de Fluorescencia por Excitación Multifotónica/métodos , Puntos Cuánticos/análisis , Animales , Ratones , Neuroimagen/métodos
16.
Opt Express ; 27(11): 15309-15317, 2019 May 27.
Artículo en Inglés | MEDLINE | ID: mdl-31163728

RESUMEN

Here, we demonstrate a polarized high-energy soliton synthesis technique for deep-brain 3-photon microscopy (3PM) excited at the 1700-nm window. Through coherent combining, we generate linearly polarized high-energy solitons whose energy is twice as high than those of each linearly polarized solitons. Due to the nonlinear origin of signals, both measured 3-photon fluorescence signal and third-harmonic signals are thus boosted by ~8 times in a tissue phantom. Using this technique, we further demonstrate 3PM of sulforhodamine 101 labeled vasculature 1600 µm in the mouse brain in vivo, which cannot be achieved by single-polarized soliton excitation.

17.
BMC Plant Biol ; 19(1): 114, 2019 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-30909879

RESUMEN

BACKGROUND: Horehound (Marrubium vulgare) is a medicinal plant whose signature bioactive compounds, marrubiin and related furanoid diterpenoid lactones, have potential applications for the treatment of cardiovascular diseases and type II diabetes. Lack of scalable plant cultivation and the complex metabolite profile of M. vulgare limit access to marrubiin via extraction from plant biomass. Knowledge of the marrubiin-biosynthetic enzymes can enable the development of metabolic engineering platforms for marrubiin production. We previously identified two diterpene synthases, MvCPS1 and MvELS, that act sequentially to form 9,13-epoxy-labd-14-ene. Conversion of 9,13-epoxy-labd-14-ene by cytochrome P450 monooxygenase (P450) enzymes can be hypothesized to facilitate key functional modification reactions in the formation of marrubiin and related compounds. RESULTS: Mining a M. vulgare leaf transcriptome database identified 95 full-length P450 candidates. Cloning and functional analysis of select P450 candidates showing high transcript abundance revealed a member of the CYP71 family, CYP71AU87, that catalyzed the hydroxylation of 9,13-epoxy-labd-14-ene to yield two isomeric products, 9,13-epoxy labd-14-ene-18-ol and 9,13-epoxy labd-14-ene-19-ol, as verified by GC-MS and NMR analysis. Additional transient Nicotiana benthamiana co-expression assays of CYP71AU87 with different diterpene synthase pairs suggested that CYP71AU87 is specific to the sequential MvCPS1 and MvELS product 9,13-epoxy-labd-14-ene. Although the P450 products were not detectable in planta, high levels of CYP71AU87 gene expression in marrubiin-accumulating tissues supported a role in the formation of marrubiin and related diterpenoids in M. vulgare. CONCLUSIONS: In a sequential reaction with the diterpene synthase pair MvCPS1 and MvELS, CYP71AU87 forms the isomeric products 9,13-epoxy labd-14-ene-18/19-ol as probable intermediates in marrubiin biosynthesis. Although its metabolic relevance in planta will necessitate further genetic studies, identification of the CYP71AU87 catalytic activity expands our knowledge of the functional landscape of plant P450 enzymes involved in specialized diterpenoid metabolism and can provide a resource for the formulation of marrubiin and related bioactive natural products.


Asunto(s)
Sistema Enzimático del Citocromo P-450/metabolismo , Diterpenos/metabolismo , Marrubium/metabolismo , Proteínas de Plantas/metabolismo , Sistema Enzimático del Citocromo P-450/química , Sistema Enzimático del Citocromo P-450/genética , Flores/genética , Flores/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Hidroxilación , Isomerismo , Marrubium/genética , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente , Plantas Medicinales/genética , Plantas Medicinales/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Especificidad por Sustrato , Nicotiana/genética
18.
J Biophotonics ; 12(6): e201800423, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-30801979

RESUMEN

Three-photon microscopy excited at the 1700-nm window (roughly covering 1600-1840 nm) is especially suitable for deep-brain imaging in living animals. To match the brain refractive index, D2 O has been exclusively used as the immersion medium. However, the hygroscopic property of D2 O leads to a decrease of transmittance of the excitation light and as a result a decrease in three-photon signals over time. Solutions such as replacing D2 O from time to time, wrapping both the objective lens and the immersion D2 O, and sealing D2 O with paraffin liquid have all been demonstrated, which add to the system complexity. Based on our recent characterization of immersion oils, we propose using silicone oil as a potential alternative to D2 O for deep-brain imaging. Excited at 1600 nm, our comparative deep-brain imaging using both D2 O and silicone oil immersion show that silicone oil immersion yields 17% higher three-photon signal in third-harmonic generation imaging within the white matter. Besides, silicone oil immersion also enables three-photon fluorescence imaging of vasculature up to 1460 µm (mechanical depth) into the mouse brain in vivo acquired at 2 seconds/frame. Together with the nonhygroscopic physical property, silicone oil is promising for long-span three-photon brain imaging excited at the 1700-nm window.


Asunto(s)
Encéfalo/diagnóstico por imagen , Microscopía de Fluorescencia por Excitación Multifotónica/métodos , Aceites de Silicona , Animales , Procesamiento de Imagen Asistido por Computador , Inmersión , Ratones , Ratones Endogámicos BALB C
19.
J Biophotonics ; 12(4): e201800360, 2019 04.
Artículo en Inglés | MEDLINE | ID: mdl-30421510

RESUMEN

Osteocytes are the most abundant cells in bone and always the focus of bone research. They are embedded in the highly scattering mineralized bone matrix. Consequently, visualizing osteocytes deep in bone with subcellular resolution poses a major challenge for in vivo bone research. Here we overcome this challenge by demonstrating 3-photon imaging of osteocytes through the intact mouse skull in vivo. Through broadband transmittance characterization, we establish that the excitation at the 1700-nm window enables the highest optical transmittance through the skull. Using label-free third-harmonic generation (THG) imaging excited at this window, we visualize osteocytes through the whole 140-µm mouse skull and 155 µm into the brain in vivo. By developing selective labeling technique for the interstitial space, we visualize the "sandwich" structure of osteocytes in their native environment. Our work provides novel imaging methodology for bone research in vivo.


Asunto(s)
Imagen Molecular , Osteocitos/citología , Cráneo/citología , Cráneo/diagnóstico por imagen , Animales , Líquidos Corporales/metabolismo , Ratones , Ratones Endogámicos BALB C
20.
Zhongguo Zhong Yao Za Zhi ; 43(5): 1034-1041, 2018 Mar.
Artículo en Chino | MEDLINE | ID: mdl-29676105

RESUMEN

The purpose of this experiment is to observe the effects of Tongbi capsule on joint lesions in rabbit with rheumatoid arthritis induced by ovalbumin and explore the mechanism in order to provide reference for clinical application of Tongbi capsule. Rheumatoid arthritis in rabbits was induced by subcutaneous injection of emulsions of ovalbumin and Freund's complete adjuvant and intra articular injection of ovalbumin. After successful modeling, 30 New Zealand rabbits with arthritis were randomly divided into model control group, the high, medium and low dose groups of Tongbi capsule (90, 45, 22.5 mg·kg⁻¹) and prednisone group (5 mg·kg⁻¹). Another six normal rabbits were used as normal control group. After 24 hours of modeling, the rabbits in Tongbi capsule groups received intragastric (i.g.) administrations of Tongbi capsule at 90, 45, 22.5 mg·kg⁻¹·d⁻¹, and the rabbits of prednisone group received i.g. administrations of prednisone at 5 mg·kg⁻¹·d⁻¹ for 2 weeks. The rabbits in normal and model groups received the same volume of distilled water at the same time. The swelling degree of rabbit knee joint and local skin temperature were observed daily. After two weeks of administration, pathological changes of rabbit knee joint were examined by magnetic resonance imaging (MRI); the morphological changes of articular cartilage and synovial membrane were observed by microscope; and the contents of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-α) in serum were detected by enzyme linked immunosorbent assay (ELISA).The results showed that 24 h after modeling, the knee joints of the rabbits were swollen, with red or dark redlocal skin, and fever, elevated local skin temperature and increased diameters of knee joints. Two weeks after modeling, the swelling of rabbit knee joints was obvious in model group; the joint cavities were filled with purulent fluid; joint synovial membranes were obviously thickened, and even joint cavities were fibrotic and cartilage surfaces showed slight defect; the surface of articular cartilage was obvious fibrosis; synovial epithelial cell proliferation was obvious and accompanied by extensive inflammatory cell infiltration; the levels of IL-1 and TNF-α were significantly higher as compared with those seen in model rabbits (P<0.05, P<0.01). After 1 and 2 weeks of administration, knee joint diameters and local skin temperatures were smaller or lower than thosein model group (P<0.05, P<0.01); The lesions of joint cartilage and synovial of all rabbits in each group were less than those in model group; IL-1 and TNF-α levels in serum were also lower than those in model group (P<0.05, P<0.01). The results reveal that high and medium doses of Tongbi capsule can suppress rheumatoid arthritis induced by ovalbumin in rabbits, reduce joint swelling, inhibit synovial epithelial and fiber hyperplasia and inflammatory cell infiltration, and alleviate articular cartilage damage. The mechanism may be associated with decreasing IL-1 and TNF-α levels in serum.


Asunto(s)
Artritis Reumatoide/tratamiento farmacológico , Cartílago Articular/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Articulaciones/efectos de los fármacos , Animales , Interleucina-1/sangre , Prednisona/farmacología , Conejos , Membrana Sinovial/efectos de los fármacos , Factor de Necrosis Tumoral alfa/sangre
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