Preparation of rhodamine-labelled tubulin and its reassembly in guard cells in leaves of Vicia faba L.

2004 ◽  
Vol 1 (3) ◽  
pp. 161-165
Author(s):  
Yu Rong ◽  
Yuan Ming ◽  
Wang Xue-Chen
Keyword(s):  
Vicia Faba ◽  
Laser Scanning ◽  
Guard Cells ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Dorsal Wall ◽  
Pig Brain ◽  
Brain Tubulin ◽  
Ventral Wall

AbstractTubulin from fresh pig brain was extracted through two cycles of polymerization and depolymerization, purified by chromatography on an ion-exchange column and labelled with 5(6)-carboxytetramethylrhodamine succinimidyl ester. We prepared 10.8 mg/ml rhodamine-labelled tubulin probe (dye/protein=0.6) and then microinjected it into living guard cells in leaves of Vicia faba L. After 5–10 min, a network of microtubules (MTs) could clearly be observed in the guard cells under a confocal laser scanning microscope. In opening guard cells, the cortical MTs radiated from the ventral wall to the dorsal wall and in closed guard cells the radial MTs were completely depolymerized into random and diffuse fragments. This shows that pig brain tubulin can easily be incorporated into MTs in guard cells, and that the dynamic behaviour of MTs can be examined directly in vivo during stomatal movement.

CPNE7 Induces Biological Dentin Sealing in a Dentin Hypersensitivity Model

2019 ◽  
Vol 98 (11) ◽  
pp. 1239-1244 ◽  
Author(s):  
S.H. Park ◽  
Y.S. Lee ◽  
D.S. Lee ◽  
J.C. Park ◽  
R. Kim ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Microscopic Analysis ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Control Group ◽  
Dentin Hypersensitivity ◽  
Dentinal Tubules ◽  
Tertiary Dentin ◽  
Dentin Formation

Dentin hypersensitivity commonly occurs due to opened dentinal tubules for many reasons. In our previous study, copine 7 (CPNE7) could induce dentin formation for an indirect pulp-capping model in vivo. This study aims to investigate the formation of tertiary dentin when CPNE7 is applied to intentionally exposed dentin with nothing over it in vivo, whether it affects microleakage of the teeth, and the penetration ability of CPNE7 molecules through dentinal tubules in vitro. Cervical dentin areas of 6 maxillary incisors of 5 beagles were exposed to a class V–like lesion, and 1 side of 3 maxillary incisors was adapted with recombinant CPNE7 protein for 5 min as the experimental group. The other side was the control group, and there was no treatment of ethylenediaminetetraacetic acid (EDTA) and CPNE7 after preparation. The defects were exposed without any restorations, and all beagles were sacrificed after 4 wk. The fluid penetration of exposed dentin areas was investigated by a microleakage-testing device and confocal laser scanning microscope. Tertiary dentin formation was confirmed with histological scanning electronic microscopic analysis. Tertiary dentin formation reduces dentinal fluid flow due to occluded tubules or discontinuity with primary or secondary dentin. The in vivo hypersensitivity model with the anterior teeth of beagle dogs showed newly formed tertiary dentin at the dentin-pulp boundary in recombinant CPNE7–treated teeth when compared with the untreated control group in histologic analysis. Scanning electronic microscopic analysis revealed occluded sites with mineral deposition of intratubular dentin. In the permeability test, the mean microleakage value of the CPNE7-treated group was significantly lower than that of the control group ( P < 0.05). The tubular penetration of rhodamine B–combined CPNE7 was confirmed under confocal laser scanning microscope. CPNE7 induces formation of tertiary dentin through shallowly exposed dentinal tubules, which decreases dentin permeability.

scholarly journals Use of Magnetic Nanoparticles to Visualize Threadlike Structures inside Lymphatic Vessels of Rats

2007 ◽  
Vol 4 (1) ◽  
pp. 77-82 ◽  
Author(s):  
Hyeon-Min Johng ◽  
Jung Sun Yoo ◽  
Tae-Jong Yoon ◽  
Hak-Soo Shin ◽  
Byung-Cheon Lee ◽  
...  
Keyword(s):  
Magnetic Nanoparticles ◽  
Laser Scanning ◽  
Lymphatic Vessels ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Scanning Microscope ◽  
Transmission Electron ◽  
Reticular Fibers ◽  
Microscope Images

A novel application of fluorescent magnetic nanoparticles was made to visualize a new tissue which had not been detectable by using simple stereomicroscopes. This unfamiliar threadlike structure inside the lymphatic vessels of rats was demonstrated in vivo by injecting nanoparticles into lymph nodes and applying magnetic fields on the collecting lymph vessels so that the nanoparticles were taken up by the threadlike structures. Confocal laser scanning microscope images of cryosectioned specimens exhibited that the nanoparticles were absorbed more strongly by the threadlike structure than by the lymphatic vessels. Further examination using a transmission electron microscope revealed that the nanoparticles had been captured between the reticular fibers in the extracellular matrix of the threadlike structures. The emerging technology of nanoparticles not only allows the extremely elusive threadlike structures to be visualized but also is expected to provide a magnetically controllable means to investigate their physiological functions.

scholarly journals In Vivo Biodistribution, Clearance, and Biocompatibility of Multiple Carbon Dots Containing Nanoparticles for Biomedical Application

Pharmaceutics ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1872
Author(s):  
Jinfeng Liao ◽  
Yuan Yao ◽  
Cheng-Hao Lee ◽  
Yongzhi Wu ◽  
Pei Li
Keyword(s):  
Carbon Dots ◽  
Laser Scanning ◽  
Biomedical Application ◽  
Confocal Laser Scanning Microscope ◽  
In Vivo Studies ◽  
Confocal Laser ◽  
In Vivo Biodistribution ◽  
Targeting Ability ◽  
Pei Nanoparticles

Current research on the use of carbon dots for various biological systems mainly focuses on the single carbon dots, while particles that contain multiple carbon dots have scarcely been investigated. Here, we assessed multiple carbon dots-crosslinked polyethyleneimine nanoparticles (CDs@PEI) for their in vivo biodistribution, clearance, biocompatibility, and cellular uptake. The in vivo studies demonstrate three unique features of the CDs@PEI nanoparticles: (1) the nanoparticles possess tumor-targeting ability with steady and prolonged retention time in the tumor region. (2) The nanoparticles show hepatobiliary excretion and are clear from the intestine in feces. (3) The nanoparticles have much better biocompatibility than the polyethyleneimine passivated single carbon dots (PEI-CD). We also found that pegylated CDs@PEI nanoparticles can be effectively taken up by the cells, which the confocal laser scanning microscope can image under different excitation wavelengths (at 405, 488, and 800 nm). These prior studies provide invaluable information and new opportunities for this new type of intrinsic photoluminescence nanoparticles in carbon dot-based biomedical applications.

scholarly journals Extraction of Three-Dimensional Information of Biological Membranous Tissue with Scanning Confocal Infrared Laser Microscope Tomography

2013 ◽  
Vol 19 (S5) ◽  
pp. 194-197 ◽  
Author(s):  
Soonwook Kwon ◽  
Se-Bum Choi ◽  
Min Gyu Park ◽  
Hyunung Yu ◽  
Seung-Woo Suh ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Near Infrared ◽  
Materials Science ◽  
Three Dimensional ◽  
Biological Tissues ◽  
Rat Aorta ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Mechanical Cutting

AbstractThe “LEXT” confocal laser scanning microscope has been used for the three-dimensional (3D) imaging of the surface of specimens, especially in materials science fields, by the penetration of near-infrared (NIR) light without mechanical cutting, deposition, or other specimen pretreatment. Noninvasive investigation of various biological tissues such as human spinal dura mater, rat aorta, and cornea without the dehydration process was successfully carried out with the “LEXT,” in order to access both surface and internal topographic images of the biological structures at a good status of the wet tissue such asin vivo, especially in measuring tissue thickness. The confocal NIR laser microscopy offers the viable means to visualize tissue architecture and its thickness in microdomain to integrate 3D images efficiently. We believe that the “LEXT” has a good application for biological researchers to study biomaterials, and it would be useful as a diagnostic tool in the near future.

3-D imaging of cells using a confocal laser scanning microscope and digital image processing

1988 ◽  
Vol 46 ◽  
pp. 96-97
Author(s):  
Thomas M. Jovin ◽  
Michel Robert-Nicoud ◽  
Donna J. Arndt-Jovin ◽  
Thorsten Schormann
Keyword(s):  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Scanning Microscope ◽  
Laser Scanning Microscope ◽  
Biochemical Processes ◽  
Adjacent Structures

Light microscopic techniques for visualizing biomolecules and biochemical processes in situ have become indispensable in studies concerning the structural organization of supramolecular assemblies in cells and of processes during the cell cycle, transformation, differentiation, and development. Confocal laser scanning microscopy offers a number of advantages for the in situ localization and quantitation of fluorescence labeled targets and probes: (i) rejection of interfering signals emanating from out-of-focus and adjacent structures, allowing the “optical sectioning” of the specimen and 3-D reconstruction without time consuming deconvolution; (ii) increased spatial resolution; (iii) electronic control of contrast and magnification; (iv) simultanous imaging of the specimen by optical phenomena based on incident, scattered, emitted, and transmitted light; and (v) simultanous use of different fluorescent probes and types of detectors.We currently use a confocal laser scanning microscope CLSM (Zeiss, Oberkochen) equipped with 3-laser excitation (u.v - visible) and confocal optics in the fluorescence mode, as well as a computer-controlled X-Y-Z scanning stage with 0.1 μ resolution.

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