scholarly journals New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains

2019 ◽  
Vol 9 (9) ◽  
pp. 218
Author(s):  
Petr Zach ◽  
Jana Mrzílková ◽  
Jan Pala ◽  
Libor Uttl ◽  
Viera Kútna ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Postmortem Brain ◽  
Confocal Laser ◽  
Original Design ◽  
Anatomical Studies ◽  
Postmortem Brain Tissue ◽  
Laboratory Results ◽  
Flow Through ◽  
Human Brains

Background: CLARITY is a method of rendering postmortem brain tissue transparent using acrylamide-based hydrogels so that this tissue could be further used for immunohistochemistry, molecular biology, or gross anatomical studies. Published papers using the CLARITY method have included studies on human brains suffering from Alzheimer’s disease using mouse spinal cords as animal models for multiple sclerosis. Methods: We modified the original design of the Chung CLARITY system by altering the electrophoretic flow-through cell, the shape of the platinum electrophoresis electrodes and their positions, as well as the cooling and recirculation system, so that it provided a greater effect and can be used in any laboratory. Results: The adapted CLARITY system is assembled from basic laboratory components, in contrast to the original design. The modified CLARITY system was tested both on rat brain stained with a rabbit polyclonal anti-Iba-1 for microglial cells and on human nucleus accumbens stained with parvalbumin and tyrosine hydroxylase for visualization of specific neurons by confocal laser scanning microscopy. Conclusions: Our design has the advantage of simplicity, functional robustness, and minimal requirement for specialized additional items for the construction of the CLARITY apparatus.

scholarly journals The Complexity of Porous Media Flow Characterized in a Microfluidic Model Based on Confocal Laser Scanning Microscopy and Micro-PIV

2020 ◽  
Author(s):  
D. A. M. de Winter ◽  
K. Weishaupt ◽  
S. Scheller ◽  
S. Frey ◽  
A. Raoof ◽  
...  
Keyword(s):  
Porous Media ◽  
Fluid Flow ◽  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Pore Scale ◽  
Micro Piv ◽  
Flow Through

Abstract In this study, the complexity of a steady-state flow through porous media is revealed using confocal laser scanning microscopy (CLSM). Micro-particle image velocimetry (micro-PIV) is applied to construct movies of colloidal particles. The calculated velocity vector fields from images are further utilized to obtain laminar flow streamlines. Fluid flow through a single straight channel is used to confirm that quantitative CLSM measurements can be conducted. Next, the coupling between the flow in a channel and the movement within an intersecting dead-end region is studied. Quantitative CLSM measurements confirm the numerically determined coupling parameter from earlier work of the authors. The fluid flow complexity is demonstrated using a porous medium consisting of a regular grid of pores in contact with a flowing fluid channel. The porous media structure was further used as the simulation domain for numerical modeling. Both the simulation, based on solving Stokes equations, and the experimental data show presence of non-trivial streamline trajectories across the pore structures. In view of the results, we argue that the hydrodynamic mixing is a combination of non-trivial streamline routing and Brownian motion by pore-scale diffusion. The results provide insight into challenges in upscaling hydrodynamic dispersion from pore scale to representative elementary volume (REV) scale. Furthermore, the successful quantitative validation of CLSM-based data from a microfluidic model fed by an electrical syringe pump provided a valuable benchmark for qualitative validation of computer simulation results. Graphic Abstract

Do we know who they are? On the identity of Pholoe (Annelida: Sigalionidae: Pholoinae) species from northern Europe

2019 ◽  
Vol 189 (1) ◽  
pp. 178-206
Author(s):  
Karin Meißner ◽  
Miriam Götting ◽  
Arne Nygren
Keyword(s):  
Coastal Waters ◽  
Laser Scanning ◽  
Taxonomic Status ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Species Relationships ◽  
Nuclear Markers ◽  
Anatomical Studies ◽  
Northern European ◽  
Species Descriptions

Abstract Pholoe species described from northern European coastal waters are studied here, based on morphological and molecular methods. We confirm the current taxonomic status of P. assimilis, P. baltica, P. inornata and P. pallida. All species descriptions are reviewed and refined, barcodes are provided allowing the identification of these species based on molecular markers. Expanding morphological methods by using confocal laser scanning microscopy has added new characters to species diagnoses and makes species identification more secure. The reliability of characters traditionally used for identification of Pholoe species is evaluated. The newly presented identification key allows the delimitation of the four species reported for European coastal waters and also includes P. minuta and P. longa. The latter have been originally described from Greenland, but were often reported from European coastal waters. The results of our studies provide new insights into the distribution of the different species. Relationships of Pholoe species from mainly northern European waters are analysed based on molecular mitochondrial and nuclear markers. We stress the need for the prospective development of a precise terminology for prostomial appendages in Pholoe by including fine anatomical studies.

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.

Vacuoles Induced in Mammalian Cells by Helicobacter Cytotoxin are Acidic Organelles

1990 ◽  
Vol 48 (3) ◽  
pp. 168-169
Author(s):  
M. H. Chestnut ◽  
C. E. Catrenich
Keyword(s):  
Acridine Orange ◽  
Mammalian Cells ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Ulcer Disease ◽  
Gastroduodenal Disease ◽  
H Pylori

Helicobacter pylori is a non-invasive, Gram-negative spiral bacterium first identified in 1983, and subsequently implicated in the pathogenesis of gastroduodenal disease including gastritis and peptic ulcer disease. Cytotoxic activity, manifested by intracytoplasmic vacuolation of mammalian cells in vitro, was identified in 55% of H. pylori strains examined. The vacuoles increase in number and size during extended incubation, resulting in vacuolar and cellular degeneration after 24 h to 48 h. Vacuolation of gastric epithelial cells is also observed in vivo during infection by H. pylori. A high molecular weight, heat labile protein is believed to be responsible for vacuolation and to significantly contribute to the development of gastroduodenal disease in humans. The mechanism by which the cytotoxin exerts its effect is unknown, as is the intracellular origin of the vacuolar membrane and contents. Acridine orange is a membrane-permeant weak base that initially accumulates in low-pH compartments. We have used acridine orange accumulation in conjunction with confocal laser scanning microscopy of toxin-treated cells to begin probing the nature and origin of these vacuoles.

Use of confocal laser scanning microscopy and a computer model to understand ink cavitation and filamentation

TAPPI Journal ◽  
2010 ◽  
Vol 9 (10) ◽  
pp. 7-15
Author(s):  
HANNA KOIVULA ◽  
DOUGLAS BOUSFIELD ◽  
MARTTI TOIVAKKA
Keyword(s):  
Laser Scanning ◽  
Confocal Laser Scanning Microscope ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Print Quality ◽  
Length Scale ◽  
Offset Printing ◽  
Significant Difference ◽  
Printing Speed

In the offset printing process, ink film splitting has an important impact on formation of ink filaments. The filament size and its distribution influence the leveling of ink and hence affect ink setting and the print quality. However, ink filaments are difficult to image due to their short lifetime and fine length scale. Due to this difficulty, limited work has been reported on the parameters that influence filament size and methods to characterize it. We imaged ink filament remains and quantified some of their characteristics by changing printing speed, ink amount, and fountain solution type. Printed samples were prepared using a laboratory printability tester with varying ink levels and operating settings. Rhodamine B dye was incorporated into fountain solutions to aid in the detection of the filaments. The prints were then imaged with a confocal laser scanning microscope (CLSM) and images were further analyzed for their surface topography. Modeling of the pressure pulses in the printing nip was included to better understand the mechanism of filament formation and the origin of filament length scale. Printing speed and ink amount changed the size distribution of the observed filament remains. There was no significant difference between fountain solutions with or without isopropyl alcohol on the observed patterns of the filament remains.

ACTIVATED SLUDGE FLOC CHARACTERIZATION BY CONFOCAL LASER SCANNING MICROSCOPY

2012 ◽  
Vol 11 (3) ◽  
pp. 669-674 ◽  
Author(s):  
Szabolcs Szilveszter ◽  
Botond Raduly ◽  
Szilard Bucs ◽  
Beata Abraham ◽  
Szabolcs Lanyi ◽  
...  
Keyword(s):  
Activated Sludge ◽  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Scanning Microscopy ◽  
Confocal Laser

Cuticular structures in the copulatory apparatus of Planorbis planorbis (Linnaeus, 1758) (Gastropoda: Pulmonata: Planorbidae)

2009 ◽  
Vol 18 (1) ◽  
pp. 11-16
Author(s):  
E.V. Soldatenko ◽  
A.A. Petrov
Keyword(s):  
Light Microscopy ◽  
Confocal Laser Scanning Microscopy ◽  
Laser Scanning ◽  
Taxonomic Status ◽  
Laser Scanning Microscopy ◽  
Confocal Laser Scanning ◽  
Confocal Laser ◽  
Copulatory Apparatus ◽  
The Family ◽  
Cuticular Structures

The morphology of the copulatory apparatus and associated cuticular structures in Planorbis planorbis was studied by light microscopy, SEM, TEM and confocal laser scanning microscopy. The significance of these cuticular structures for the taxonomic status of the species and for the systematics of the family Planorbidae in general is discussed.

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