scholarly journals An integrated system for synchronous culture of animal cells under controlled conditions

BioTechniques ◽  
2016 ◽  
Vol 61 (3) ◽  
Author(s):  
Elena Mendoza-Pérez ◽  
Vanessa Hernández ◽  
Laura A. Palomares ◽  
José A. Serrato
Keyword(s):  
Integrated System ◽  
Synchronous Culture ◽  
Animal Cells ◽  
Controlled Conditions

scholarly journals Contractile ring constriction and septation in fission yeast are integrated mutually stabilizing processes

2021 ◽  
Author(s):  
Sathish Thiyagarajan ◽  
Zachary A McDargh ◽  
Shuyuan Wang ◽  
Ben O'Shaughnessy
Keyword(s):  
Cell Wall ◽  
Fission Yeast ◽  
Growth Ring ◽  
Integrated System ◽  
Animal Cells ◽  
Contractile Ring ◽  
Cell Wall Growth ◽  
Ring Constriction ◽  
Wall Growth ◽  
In Cells

In common with other cellular machineries, the actomyosin contractile ring that divides cells during cytokinesis does not operate in isolation. Contractile rings in animal cells interact with contiguous actomyosin cortex, while ring constriction in many cell-walled organisms couples tightly to cell wall growth. In fission yeast, a septum grows in the wake of the constricting ring, ensuring cytokinesis leaves two daughter cells fully enclosed by cell wall. Here we mathematical modeled the integrated constriction-septation system in fission yeast, with a kinetic growth model evolving the 3D septum shape coupled to a molecularly explicit simulation of the contractile ring highly constrained by experimental data. Simulations revealed influences in both directions, stabilizing the ring-septum system as a whole. By providing a smooth circular anchoring surface for the ring, the inner septum leading edge stabilized ring organization and tension production; by mechanically regulating septum circularity and in-plane growth, ring tension stabilized septum growth and shape. Genetic or pharmacological perturbation of either subsystem destabilized this delicate balance, precipitating uncontrolled positive feedback with disastrous morphological and functional consequences. Thus, high curvature septum irregularities triggered bridging instabilities, in which contractile ring segments became unanchored. Bridging abolished the local tension-mediated septum shape regulation, exacerbating the irregularity in a mutually destabilizing runaway process. Our model explains a number of previously mysterious experimental observations, including unanchoring of ring segments observed in cells with mutations in the septum-growing β-glucan synthases, and irregular septa in cells with mutations in the contractile ring myosin-II Myo2. Thus, the contractile ring and cell wall growth cellular machineries operate as a single integrated system, whose stability relies on mutual regulation by the two subsystems.

Surface Structure of Nucleated Animal Cells: Carbon Replicas

1967 ◽  
Vol 25 ◽  
pp. 120-121
Author(s):  
Robert M. Glaeser ◽  
Thea B. Scott
Keyword(s):  
Cell Surface ◽  
Surface Structure ◽  
Particle Diameter ◽  
Cellular Functions ◽  
Animal Cells ◽  
Replica Technique ◽  
Carbon Replica ◽  
Characteristic Particle ◽  
Cell Surface Structure ◽  
Carbon Replicas

The carbon-replica technique can be used to obtain information about cell-surface structure that cannot ordinarily be obtained by thin-section techniques. Mammalian erythrocytes have been studied by the replica technique and they appear to be characterized by a pebbly or “plaqued“ surface texture. The characteristic “particle” diameter is about 200 Å to 400 Å. We have now extended our observations on cell-surface structure to chicken and frog erythrocytes, which possess a broad range of cellular functions, and to normal rat lymphocytes and mouse ascites tumor cells, which are capable of cell division. In these experiments fresh cells were washed in Eagle's Minimum Essential Medium Salt Solution (for suspension cultures) and one volume of a 10% cell suspension was added to one volume of 2% OsO4 or 5% gluteraldehyde in 0.067 M phosphate buffer, pH 7.3. Carbon replicas were obtained by a technique similar to that employed by Glaeser et al. Figure 1 shows an electron micrograph of a carbon replica made from a chicken erythrocyte, and Figure 2 shows an enlarged portion of the same cell.

Development of a Rapid Survey Method of Sampling and Analysis for Asbestos in Ambient Air

1972 ◽  
Vol 30 ◽  
pp. 630-631 ◽  
Author(s):  
R. E. Heffelfinger ◽  
C. W. Melton ◽  
D. L. Kiefer ◽  
W. M. Henry ◽  
R. J. Thompson
Keyword(s):  
Neutron Activation ◽  
Ambient Air ◽  
Survey Method ◽  
Potential Hazard ◽  
Transmission Microscopy ◽  
Comparative Analyses ◽  
Asbestos Fibers ◽  
Controlled Conditions ◽  
Unequivocal Identification ◽  
Identification And Quantification

A methodology has been developed and demonstrated which is capable of determining total amounts of asbestos fibers and fibrils in air ranging from as low as fractional nanograms per cubic meter (ng/m3) of air to several micrograms/m3. The method involves the collection of samples on an absolute filter and provides an unequivocal identification and quantification of the total asbestos contents including fibrils in the collected samples.The developed method depends on the trituration under controlled conditions to reduce the fibers to fibrils, separation of the asbestos fibrils from other collected air particulates (beneficiation), and the use of transmission microscopy for identification and quantification. Its validity has been tested by comparative analyses by neutron activation techniques. It can supply the data needed to set emissions criteria and to serve as a basis for assessing the potential hazard for asbestos pollution to the populace.

Nuclear Envelope Dynamics During Mitosis

1988 ◽  
Vol 46 ◽  
pp. 224-225
Author(s):  
Brian Burke
Keyword(s):  
Nuclear Envelope ◽  
Membrane Vesicles ◽  
Nuclear Pore ◽  
Nuclear Pore Complexes ◽  
Animal Cells ◽  
Interphase Chromosome ◽  
Lamin B ◽  
Chromosome Architecture ◽  
Complex Membrane ◽  
Pore Complexes

The nuclear envelope is a complex membrane structure that forms the boundary of the nuclear compartment in eukaryotes. It regulates the passage of macromolecules between the two compartments and may be important for organizing interphase chromosome architecture. In interphase animal cells it forms a remarkably stable structure consisting of a double membrane ouerlying a protein meshwork or lamina and penetrated by nuclear pore complexes. The latter form the channels for nucleocytoplasmic exchange of macromolecules, At the onset of mitosis, however, it rapidly disassembles, the membranes fragment to yield small vesicles and the lamina, which is composed of predominantly three polypeptides, lamins R, B and C (MW approx. 74, 68 and 65 kDa respectiuely), breaks down. Lamins B and C are dispersed as monomers throughout the mitotic cytoplasm, while lamin B remains associated with the nuclear membrane vesicles.

Localization of Adenosine Triphosphatase Activity in the Phleom of Nicotiana Tabacum

1972 ◽  
Vol 30 ◽  
pp. 230-231
Author(s):  
James Cronshaw ◽  
Jamison E. Gilder
Keyword(s):  
Plasma Membrane ◽  
Atpase Activity ◽  
Adenosine Triphosphatase ◽  
Higher Plants ◽  
High Surface ◽  
Root Tip ◽  
Animal Cells ◽  
Physiological Processes ◽  
Tip Cells ◽  
Atpase Localization

Adenosine triphosphatase (ATPase) activity has been shown to be associated with numerous physiological processes in both plants and animal cells. Biochemical studies have shown that in higher plants ATPase activity is high in cell wall preparations and is associated with the plasma membrane, nuclei, mitochondria, chloroplasts and lysosomes. However, there have been only a few ATPase localization studies of higher plants at the electron microscope level. Poux (1967) demonstrated ATPase activity associated with most cellular organelles in the protoderm cells of Cucumis roots. Hall (1971) has demonstrated ATPase activity in root tip cells of Zea mays. There was high surface activity largely associated with the plasma membrane and plasmodesmata. ATPase activity was also demonstrated in mitochondria, dictyosomes, endoplasmic reticulum and plastids.

FR-IR Molecular Microanalysis System

1990 ◽  
Vol 48 (2) ◽  
pp. 270-271
Author(s):  
John A. Reffner ◽  
William T. Wihlborg
Keyword(s):  
Fourier Transform ◽  
Fourier Transform Infrared ◽  
Integrated System ◽  
Morphological Examination ◽  
Fully Integrated ◽  
Ir Microscopy ◽  
Normal Functions ◽  
Infrared Microspectroscopy ◽  
Infrared Spectral ◽  
Ft Ir

The IRμs™ is the first fully integrated system for Fourier transform infrared (FT-IR) microscopy. FT-IR microscopy combines light microscopy for morphological examination with infrared spectroscopy for chemical identification of microscopic samples or domains. Because the IRμs system is a new tool for molecular microanalysis, its optical, mechanical and system design are described to illustrate the state of development of molecular microanalysis. Applications of infrared microspectroscopy are reviewed by Messerschmidt and Harthcock.Infrared spectral analysis of microscopic samples is not a new idea, it dates back to 1949, with the first commercial instrument being offered by Perkin-Elmer Co. Inc. in 1953. These early efforts showed promise but failed the test of practically. It was not until the advances in computer science were applied did infrared microspectroscopy emerge as a useful technique. Microscopes designed as accessories for Fourier transform infrared spectrometers have been commercially available since 1983. These accessory microscopes provide the best means for analytical spectroscopists to analyze microscopic samples, while not interfering with the FT-IR spectrometer’s normal functions.

Structural aspects of tracheary element differentiation in suspension cultures of Zinnia elegans

1989 ◽  
Vol 47 ◽  
pp. 768-769
Author(s):  
C. H. Haigler ◽  
A. W. Roberts
Keyword(s):  
Tracheary Element ◽  
Vesicle Fusion ◽  
Zinnia Elegans ◽  
Cellulose Synthesis ◽  
Tracheary Elements ◽  
Animal Cells ◽  
Mesophyll Cells ◽  
Fixation Techniques ◽  
Localized Patterns ◽  
Structural Aspects

Tracheary elements, the water-conducting cells in plants, are characterized by their reinforced walls that became thickened in localized patterns during differentiation (Fig. 1). The synthesis of this localized wall involves abundant secretion of Golgi vesicles that export preformed matrix polysaccharides and putative proteins involved in cellulose synthesis. Since the cells are not growing, some kind of endocytotic process must also occur. Many researchers have commented on where exocytosis occurs in relation to the thickenings (for example, see), but they based their interpretations on chemical fixation techniques that are not likely to provide reliable information about rapid processes such as vesicle fusion. We have used rapid freezing to more accurately assess patterns of vesicle fusion in tracheary elements. We have also determined the localization of calcium, which is known to regulate vesicle fusion in plant and animal cells.Mesophyll cells were obtained from immature first leaves of Zinnia elegans var. Envy (Park Seed Co., Greenwood, S.C.) and cultured as described previously with the following exceptions: (a) concentration of benzylaminopurine in the culture medium was reduced to 0.2 mg/l and myoinositol was eliminated; and (b) 1.75ml cultures were incubated in 22 x 90mm shell vials with 112rpm rotary shaking. Cells that were actively involved in differentiation were harvested and frozen in solidifying Freon as described previously. Fractures occurred preferentially at the cell/planchet interface, which allowed us to find some excellently-preserved cells in the replicas. Other differentiating cells were incubated for 20-30 min in 10(μM CTC (Sigma), an antibiotic that fluoresces in the presence of membrane-sequestered calcium. They were observed in an Olympus BH-2 microscope equipped for epi-fluorescence (violet filter package and additional Zeiss KP560 barrier filter to block chlorophyll autofluorescence).

Three-Dimensional reconstruction of isolated centrosomes by automated electron tomography

1994 ◽  
Vol 52 ◽  
pp. 310-311
Author(s):  
M.B. Braunfeld ◽  
M. Moritz ◽  
B.M. Alberts ◽  
J.W. Sedat ◽  
D.A. Agard
Keyword(s):  
Electron Tomography ◽  
Three Dimensional ◽  
Vital Role ◽  
Complex Nature ◽  
Animal Cells ◽  
Microtubule Organizing Center ◽  
Nucleation Sites ◽  
Dimensional Reconstruction ◽  
Organizing Center ◽  
Resolution Model

In animal cells, the centrosome functions as the primary microtubule organizing center (MTOC). As such the centrosome plays a vital role in determining a cell's shape, migration, and perhaps most importantly, its division. Despite the obvious importance of this organelle little is known about centrosomal regulation, duplication, or how it nucleates microtubules. Furthermore, no high resolution model for centrosomal structure exists.We have used automated electron tomography, and reconstruction techniques in an attempt to better understand the complex nature of the centrosome. Additionally we hope to identify nucleation sites for microtubule growth.Centrosomes were isolated from early Drosophila embryos. Briefly, after large organelles and debris from homogenized embryos were pelleted, the resulting supernatant was separated on a sucrose velocity gradient. Fractions were collected and assayed for centrosome-mediated microtubule -nucleating activity by incubating with fluorescently-labeled tubulin subunits. The resulting microtubule asters were then spun onto coverslips and viewed by fluorescence microscopy.

Fate of sperm membrane in fertilized ova

1993 ◽  
Vol 51 ◽  
pp. 40-41
Author(s):  
Frank J. Longo
Keyword(s):  
Electron Microscopy ◽  
Electrical Activity ◽  
Sea Urchins ◽  
Morphological Changes ◽  
Integrated System ◽  
Electrophysiological Recording ◽  
Experimental Conditions ◽  
The Status ◽  
Sperm Membrane ◽  
Temporal And Spatial

Measurement of the egg's electrical activity, the fertilization potential or the activation current (in voltage clamped eggs), provides a means of detecting the earliest perceivable response of the egg to the fertilizing sperm. By using the electrical physiological record as a “real time” indicator of the instant of electrical continuity between the gametes, eggs can be inseminated with sperm at lower, more physiological densities, thereby assuring that only one sperm interacts with the egg. Integrating techniques of intracellular electrophysiological recording, video-imaging, and electron microscopy, we are able to identify the fertilizing sperm precisely and correlate the status of gamete organelles with the first indication (fertilization potential/activation current) of the egg's response to the attached sperm. Hence, this integrated system provides improved temporal and spatial resolution of morphological changes at the site of gamete interaction, under a variety of experimental conditions. Using these integrated techniques, we have investigated when sperm-egg plasma membrane fusion occurs in sea urchins with respect to the onset of the egg's change in electrical activity.

Animal Cells as Bioreactors

1994 ◽  
Author(s):  
Terence Cartwright
Keyword(s):  
Animal Cells
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