scholarly journals Actin Filament Disruption Alters Phragmoplast Microtubule Dynamics during the Initial Phase of Plant Cytokinesis

2020 ◽  
Vol 61 (3) ◽  
pp. 445-456 ◽  
Author(s):  
Keisho Maeda ◽  
Michiko Sasabe ◽  
Shigeru Hanamata ◽  
Yasunori Machida ◽  
Seiichiro Hasezawa ◽  
...  
Keyword(s):  
Actin Filaments ◽  
Initial Phase ◽  
Laser Scanning ◽  
Actin Polymerization ◽  
Late Phase ◽  
Membrane Vesicles ◽  
Cell Plate ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Plant Cytokinesis

Abstract Plant growth and development relies on the accurate positioning of the cell plate between dividing cells during cytokinesis. The cell plate is synthetized by a specialized structure called the phragmoplast, which contains bipolar microtubules that polymerize to form a framework with the plus ends at or near the division site. This allows the transport of Golgi-derived vesicles toward the plus ends to form and expand the cell plate. Actin filaments play important roles in cell plate expansion and guidance in plant cytokinesis at the late phase, but whether they are involved at the early phase is unknown. To investigate this further, we disrupted the actin filaments in cell cycle-synchronized tobacco BY-2 cells with latrunculin B (LatB), an actin polymerization inhibitor. We observed the cells under a transmission electron microscope or a spinning-disk confocal laser scanning microscope. We found that disruption of actin filaments by LatB caused the membrane vesicles at the equatorial plane of the cell plate to be dispersed rather than form clusters as they did in the untreated cells. The midzone constriction of phragmoplast microtubules also was perturbed in LatB-treated cells. The live cell imaging and kymograph analysis showed that disruption of actin filaments also changed the accumulation timing of NACK1 kinesin, which plays a crucial role in cell plate expansion. This suggests that there are two functionally different types of microtubules in the phragmoplast. Together, our results show that actin filaments regulate phragmoplast microtubules at the initial phase of plant cytokinesis.

Distribution of Membranes and the Cytoskeleton During Cell Plate Formation in Pollen Mother Cells of Tradescantia

1991 ◽  
Vol 100 (4) ◽  
pp. 717-728 ◽  
Author(s):  
CHRISTEL R. SCHOPFER ◽  
PETER K. HEPLER
Keyword(s):  
Laser Scanning ◽  
Polarized Light ◽  
Cell Plate ◽  
Confocal Laser Scanning Microscope ◽  
Temporal Organization ◽  
Ion Concentration ◽  
Confocal Laser ◽  
Surface Binding ◽  
Pollen Mother Cells ◽  
Mother Cells

The cellular pattern and distribution of membranes have been analyzed during cytokinesis in pollen mother cells of Tradescantia and compared with those of actin microfilaments (MFs) and microtubules (MTs). Membranes have been stained with DiOC6(3) and MFs with rhodamine-labeled phalloidin (RP); analysis has been carried out on the confocal laser scanning microscope. MTs have been visualized as birefringent elements in the polarized light microscope. The results show that when the interzone first appears in mid anaphase it contains an even distribution of membranes. However, by late anaphase these elements have been cleared away, leaving the interzone largely devoid of DiOC6(3)-positive material. MTs are found throughout this zone, while MFs appear in two non-overlapping sets on both sides of the cell equator. Thereafter membrane elements reappear in the interzone, but only along the equatorial line of the forming cell plate. Presumably these equatorial elements are composed of endoplasmic reticulum and Golgi vesicles, since the larger organelles, including amyloplasts and mitochondria, are excluded from the phragmoplast. MFs, like MTs, arrange preferentially normal to the cell plate, forming a dense array on both sides, but being absent from the zone occupied by the membranes. By contrast, the parallel set of MTs, while excluding larger organelles from the phragmoplast, intermingle with the membrane elements in the cell equator. As cytokinesis proceeds membranes continue to concentrate on the cell plate as indicated by its marked increase in staining with DiOC6(3). From a consideration of spatial and temporal organization of the phragmoplast components it is reasonable to suggest that both cytoskeletal components participate in the aggregation of vesicles that give rise to the cell plate. Membranes, on the other hand, through the provision of surface binding sites and/or through the regulation of the cytoplasmic calcium ion concentration, might be involved in the assembly and stabilization of the cytoskeleton.

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.

3-Dimensional immunolabeling and in situ hybridization detection using fluorescence photooxidation and intermediate-voltage Electron Microscopy

1994 ◽  
Vol 52 ◽  
pp. 164-165
Author(s):  
Thomas J. Deerinck ◽  
Maryann E. Martone ◽  
Varda Lev-Ram ◽  
David P. L. Green ◽  
Roger Y. Tsien ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Reactive Oxygen ◽  
Confocal Laser Scanning Microscope ◽  
Fluorescent Dye ◽  
Confocal Laser ◽  
3 Dimensional ◽  
Voltage Electron ◽  
Dimensional Distribution ◽  
Electron Microscopes ◽  
New Generation

The confocal laser scanning microscope has become a powerful tool in the study of the 3-dimensional distribution of proteins and specific nucleic acid sequences in cells and tissues. This is also proving to be true for a new generation of high contrast intermediate voltage electron microscopes (IVEM). Until recently, the number of labeling techniques that could be employed to allow examination of the same sample with both confocal and IVEM was rather limited. One method that can be used to take full advantage of these two technologies is fluorescence photooxidation. Specimens are labeled by a fluorescent dye and viewed with confocal microscopy followed by fluorescence photooxidation of diaminobenzidine (DAB). In this technique, a fluorescent dye is used to photooxidize DAB into an osmiophilic reaction product that can be subsequently visualized with the electron microscope. The precise reaction mechanism by which the photooxidation occurs is not known but evidence suggests that the radiationless transfer of energy from the excited-state dye molecule undergoing the phenomenon of intersystem crossing leads to the formation of reactive oxygen species such as singlet oxygen. It is this reactive oxygen that is likely crucial in the photooxidation of DAB.

A confocal laser scanning microscope for fluorescence and reflection at video rates

1989 ◽  
Vol 47 ◽  
pp. 134-135
Author(s):  
P.M. Houpt ◽  
A. Draaijer
Keyword(s):  
Light Source ◽  
Laser Scanning ◽  
Focal Point ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Point Light Source ◽  
Volume Of Interest ◽  
Ion Laser ◽  
Point Light ◽  
Point Detector

In confocal microscopy, the object is scanned by the coinciding focal points (confocal) of a point light source and a point detector both focused on a certain plane in the object. Only light coming from the focal point is detected and, even more important, out-of-focus light is rejected.This makes it possible to slice up optically the ‘volume of interest’ in the object by moving it axially while scanning the focused point light source (X-Y) laterally. The successive confocal sections can be stored in a computer and used to reconstruct the object in a 3D image display.The instrument described is able to scan the object laterally with an Ar ion laser (488 nm) at video rates. The image of one confocal section of an object can be displayed within 40 milliseconds (1000 х 1000 pixels). The time to record the total information within the ‘volume of interest’ normally depends on the number of slices needed to cover it, but rarely exceeds a few seconds.

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.

scholarly journals Nonlinear absorption and scattering of a single plasmonic nanostructure characterized by x-scan technique

2019 ◽  
Vol 10 ◽  
pp. 2182-2191 ◽  
Author(s):  
Tushar C Jagadale ◽  
Dhanya S Murali ◽  
Shi-Wei Chu
Keyword(s):  
Laser Scanning ◽  
Detection System ◽  
Nonlinear Behavior ◽  
Optical Nonlinearity ◽  
Research Area ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Thin Sample ◽  
Channel Detection ◽  
Novel Method

Nonlinear nanoplasmonics is a largely unexplored research area that paves the way for many exciting applications, such as nanolasers, nanoantennas, and nanomodulators. In the field of nonlinear nanoplasmonics, it is highly desirable to characterize the nonlinearity of the optical absorption and scattering of single nanostructures. Currently, the common method to quantify optical nonlinearity is the z-scan technique, which yields real and imaginary parts of the permittivity by moving a thin sample with a laser beam. However, z-scan typically works with thin films, and thus acquires nonlinear responses from ensembles of nanostructures, not from single ones. In this work, we present an x-scan technique that is based on a confocal laser scanning microscope equipped with forward and backward detectors. The two-channel detection offers the simultaneous quantification for the nonlinear behavior of scattering, absorption and total attenuation by a single nanostructure. At low excitation intensities, both scattering and absorption responses are linear, thus confirming the linearity of the detection system. At high excitation intensities, we found that the nonlinear response can be derived directly from the point spread function of the x-scan images. Exceptionally large nonlinearities of both scattering and absorption are unraveled simultaneously for the first time. The present study not only provides a novel method for characterizing nonlinearity of a single nanostructure, but also reports surprisingly large plasmonic nonlinearities.

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