C-Microtubules in Isolated Mitotic Spindles

1971 ◽  
Vol 9 (3) ◽  
pp. 603-619
Author(s):  
W. D. COHEN ◽  
T. GOTTLIEB
Keyword(s):  
Cross Section ◽  
Sea Urchin ◽  
Cross Sections ◽  
Metaphase Chromosome ◽  
Inverse Relationship ◽  
Mitotic Spindles ◽  
Cylindrical Structure ◽  
Arbacia Punctulata ◽  
Alternate Possibility

Microtubules with incomplete cylindrical structure are present in isolated mitotic spindles of the sea urchin, Arbacia punctulata. In cross-section they appear C-shaped, and are thus similar to the ‘C-microtubules’ or ‘C-filaments’ observed previously in other systems. The C-microtubules are not uniformly distributed within isolated spindles, but are typically numerous in the interzonal region of anaphase spindles and in the metaphase chromosome ‘plate’. In chromosome-to-pole regions they are seen much less frequently, and microtubules with the usual O-configuration predominate. Counts of C- and O-microtubules in anaphase spindle cross-sections of known location show an inverse relationship between the number of C-microtubules present and the total number of microtubules present. The observations suggest that the C-microtubules are not simple artifacts of fixation or isolation, but rather may represent a stage of microtubule disassembly which occurs in the interzone during isolation or during anaphase in vivo. The alternate possibility of assembly is not excluded, however. The significance of C-microtubules is further discussed with respect to their occurrence in other systems, and to potential differences between mitotic microtubules.

scholarly journals Calcium-labile mitotic spindles isolated from sea urchin eggs (Lytechinus variegatus).

1980 ◽  
Vol 86 (2) ◽  
pp. 355-365 ◽  
Author(s):  
E D Salmon ◽  
R R Segall
Keyword(s):  
Sea Urchin ◽  
Calcium Ions ◽  
Divalent Cations ◽  
Lytechinus Variegatus ◽  
Mitotic Spindles ◽  
Mitotic Apparatus ◽  
Spindle Microtubules ◽  
Membrane Components ◽  
Low Ionic Strength

We isolated calcium-labile mitotic spindles from eggs of the sea urchin Lytechinus variegatus, using a low ionic strength, EGTA lysis buffer that contined 5.0 mM EGTA, 0.5 mM MgCl2, 10-50 mM PIPES, pH 6.8, with 1% Nonidet P-40 (detergent) and 20-25% glycerol. Isolated spindles were stored in EGTA buffer with 50% glycerol for 5-6 wk without deterioration. The isolated spindles were composed primarily of microtubules with the chromosomes attached. No membranes were seen. Isolated spindles, perfused with EGTA buffer to remove the detergent and glycerol, had essentially the same birefringent retardation (BR) as spindles in vivo at the same mitotic stage. Even in the absence of glycerol and exogenous tubulin, the isolated spindles were relatively stable in the EGTA buffer: BR decayed slowly to about half the initial value within 30-45 min. However, both the rate and extent of BR decay increased with concentrations of Ca2+ above 0.2-0.5 muM as assayed using Ca-EGTA buffers (0.2 mM EGTA, 0.5 mM MgCl2, 50 mM PIPES, pH 6.8, plus various amounts of CaCl2). Microtubules depolymerized almost completely in < 6 min at Ca2+ concentrations of 2 muM and within several seconds at 10 muM Ca2+. Of several divalent cations tested, only Sr2+ caused comparable changes in BR. The absence of membranes in the isolated spindles appeared to be associated with a lack of calcium-sequestering ability. Our results suggest that calcium ions play an important role in the depolymerization of spindle microtubules and that membrane components may function within the mitotic apparatus of living cells to sequester and release calcium ions during mitosis.

An Estimate of the Amount of Microtubule Protein in the Isolated Mitotic Apparatus

1970 ◽  
Vol 6 (1) ◽  
pp. 159-176
Author(s):  
W. D. COHEN ◽  
L. I. REBHUN
Keyword(s):  
Electron Microscopy ◽  
Sea Urchin ◽  
Cross Sections ◽  
Mitotic Apparatus ◽  
Arbacia Punctulata ◽  
Molecular Weights ◽  
Cellular Factors ◽  
Microtubule Protein ◽  
Specific Orientation ◽  
Existing Data

The microtubule content of the isolated mitotic apparatus of sea-urchin eggs (Arbacia punctulata has been investigated by electron microscopy. Cross-sections were made through asters or spindles of flat-embedded mitotic apparatuses of known mitotic stage and specific orientation in the block. Cross-sections between chromosomes and poles of five metaphase half-spindles revealed approximately 2000-2300 sectioned microtubules. The number was somewhat higher in three anaphase half-spindles examined, approximately 2400-2600. A method was devised for calculating the total number of microtubules in an aster, based upon the number of microtubules appearing in cross-sections. Application of this method to selected mitotic apparatuses enabled calculation of the total number of microtubules in metaphase mitotic apparatuses of average dimensions. Using a 13-protofilament model of the microtubule and existing data on possible monomer sizes and molecular weights, the total amount of microtubule protein in the isolated mitotic apparatus was calculated. The values obtained are in the range of about 1-2 x 10-8 mg microtubule protein per isolated mitotic apparatus. These values are close to those reported for the 4-5s protein of the isolated mitotic apparatus, but are considerably lower than the amount of 22s protein. The results are discussed with respect to cellular factors which determine microtubule number, and the possible sources and origin of mitotic microtubule protein.

Electron Irradiation Effects in Polyoxymethylene Crystals Grown Inside Trioxane Crystals

1971 ◽  
Vol 29 ◽  
pp. 78-79
Author(s):  
J. P. Colson ◽  
D. H. Reneker
Keyword(s):  
Cross Section ◽  
Cross Sections ◽  
Detailed Examination ◽  
The Other ◽  
Electron Micrograph ◽  
Irradiation Effects ◽  
Threefold Axis ◽  
Typical Sample ◽  
Polymer Crystals ◽  
Chain Axis

Polyoxymethylene (POM) crystals grow inside trioxane crystals which have been irradiated and heated to a temperature slightly below their melting point. Figure 1 shows a low magnification electron micrograph of a group of such POM crystals. Detailed examination at higher magnification showed that three distinct types of POM crystals grew in a typical sample. The three types of POM crystals were distinguished by the direction that the polymer chain axis in each crystal made with respect to the threefold axis of the trioxane crystal. These polyoxymethylene crystals were described previously.At low magnifications the three types of polymer crystals appeared as slender rods. One type had a hexagonal cross section and the other two types had rectangular cross sections, that is, they were ribbonlike.

A quantitative approach to inner shell losses

1978 ◽  
Vol 36 (1) ◽  
pp. 526-527 ◽  
Author(s):  
R.D. Leapman ◽  
P. Rez ◽  
D.F. Mayers
Keyword(s):  
Energy Transfer ◽  
Cross Section ◽  
Cross Sections ◽  
Accurate Determination ◽  
Background Intensity ◽  
Core Excitation ◽  
Quantitative Basis ◽  
Cross Section Differential ◽  
Bound Electrons

Microanalysis by EELS has been developing rapidly and though the general form of the spectrum is now understood there is a need to put the technique on a more quantitative basis (1,2). Certain aspects important for microanalysis include: (i) accurate determination of the partial cross sections, σx(α,ΔE) for core excitation when scattering lies inside collection angle a and energy range ΔE above the edge, (ii) behavior of the background intensity due to excitation of less strongly bound electrons, necessary for extrapolation beneath the signal of interest, (iii) departures from the simple hydrogenic K-edge seen in L and M losses, effecting σx and complicating microanalysis. Such problems might be approached empirically but here we describe how computation can elucidate the spectrum shape.The inelastic cross section differential with respect to energy transfer E and momentum transfer q for electrons of energy E0 and velocity v can be written as

Oxygen K near edge structures studied with multiple scattering calculations

1988 ◽  
Vol 46 ◽  
pp. 504-505
Author(s):  
Xudong Weng ◽  
Peter Rez
Keyword(s):  
Cross Section ◽  
Cross Sections ◽  
Partial Cross Section ◽  
Energy Window ◽  
Edge Structure ◽  
Fine Structures ◽  
Hydrogenic Model ◽  
Electron Energy Loss ◽  
Quantitative Chemical

In electron energy loss spectroscopy, quantitative chemical microanalysis is performed by comparison of the intensity under a specific inner shell edge with the corresponding partial cross section. There are two commonly used models for calculations of atomic partial cross sections, the hydrogenic model and the Hartree-Slater model. Partial cross sections could also be measured from standards of known compositions. These partial cross sections are complicated by variations in the edge shapes, such as the near edge structure (ELNES) and extended fine structures (ELEXFS). The role of these solid state effects in the partial cross sections, and the transferability of the partial cross sections from material to material, has yet to be fully explored. In this work, we consider the oxygen K edge in several oxides as oxygen is present in many materials. Since the energy window of interest is in the range of 20-100 eV, we limit ourselves to the near edge structures.

Absolute inner-shell cross section determination for EELS analysis

1988 ◽  
Vol 46 ◽  
pp. 534-535
Author(s):  
P.A. Crozier
Keyword(s):  
Cross Section ◽  
Cross Sections ◽  
Absolute Cross Section ◽  
Specimen Thickness ◽  
Mass Thickness ◽  
Scattering Cross ◽  
Before And After ◽  
Estimated Error ◽  
Scattering Cross Sections ◽  
Electron Microscopist

Absolute inelastic scattering cross sections or mean free paths are often used in EELS analysis for determining elemental concentrations and specimen thickness. In most instances, theoretical values must be used because there have been few attempts to determine experimental scattering cross sections from solids under the conditions of interest to electron microscopist. In addition to providing data for spectral quantitation, absolute cross section measurements yields useful information on many of the approximations which are frequently involved in EELS analysis procedures. In this paper, experimental cross sections are presented for some inner-shell edges of Al, Cu, Ag and Au.Uniform thin films of the previously mentioned materials were prepared by vacuum evaporation onto microscope cover slips. The cover slips were weighed before and after evaporation to determine the mass thickness of the films. The estimated error in this method of determining mass thickness was ±7 x 107g/cm2. The films were floated off in water and mounted on Cu grids.

3-D reconstruction and analysis of mammalian mitotic spindle structure

1992 ◽  
Vol 50 (1) ◽  
pp. 578-579
Author(s):  
Kent McDonald ◽  
David Mastronarde ◽  
Rubai Ding ◽  
Eileen O'Toole ◽  
J. Richard McIntosh
Keyword(s):  
Cross Sections ◽  
Mitotic Spindle ◽  
Experimental Studies ◽  
Video Camera ◽  
Three Dimensions ◽  
Mitotic Spindles ◽  
Black And White ◽  
Reconstruction Methods ◽  
Spindle Structure ◽  
Tissue Culture Line

Mammalian spindles are generally large and may contain over a thousand microtubules (MTs). For this reason they are difficult to reconstruct in three dimensions and many researchers have chosen to study the smaller and simpler spindles of lower eukaryotes. Nevertheless, the mammalian spindle is used for many experimental studies and it would be useful to know its detailed structure.We have been using serial cross sections and computer reconstruction methods to analyze MT distributions in mitotic spindles of PtK cells, a mammalian tissue culture line. Images from EM negatives are digtized on a light box by a Dage MTI video camera containing a black and white Saticon tube. The signal is digitized by a Parallax 1280 graphics device in a MicroVax III computer. Microtubules are digitized at a magnification such that each is 10-12 pixels in diameter.

A technique for preparing semiconductor cross sections for both TEM and SEM analysis

1989 ◽  
Vol 47 ◽  
pp. 712-713
Author(s):  
Stanley J. Klepeis ◽  
J.P. Benedict ◽  
R.M Anderson
Keyword(s):  
Sample Preparation ◽  
Cross Section ◽  
Cross Sections ◽  
Sem Analysis ◽  
Preparation Technique ◽  
Ion Milling ◽  
Tem Analysis ◽  
Polished Cross Section ◽  
Area Of Interest ◽  
Polished Side

The ability to prepare a cross-section of a specific semiconductor structure for both SEM and TEM analysis is vital in characterizing the smaller, more complex devices that are now being designed and manufactured. In the past, a unique sample was prepared for either SEM or TEM analysis of a structure. In choosing to do SEM, valuable and unique information was lost to TEM analysis. An alternative, the SEM examination of thinned TEM samples, was frequently made difficult by topographical artifacts introduced by mechanical polishing and lengthy ion-milling. Thus, the need to produce a TEM sample from a unique,cross-sectioned SEM sample has produced this sample preparation technique.The technique is divided into an SEM and a TEM sample preparation phase. The first four steps in the SEM phase: bulk reduction, cleaning, gluing and trimming produces a reinforced sample with the area of interest in the center of the sample. This sample is then mounted on a special SEM stud. The stud is inserted into an L-shaped holder and this holder is attached to the Klepeis polisher (see figs. 1 and 2). An SEM cross-section of the sample is then prepared by mechanically polishing the sample to the area of interest using the Klepeis polisher. The polished cross-section is cleaned and the SEM stud with the attached sample, is removed from the L-shaped holder. The stud is then inserted into the ion-miller and the sample is briefly milled (less than 2 minutes) on the polished side. The sample on the stud may then be carbon coated and placed in the SEM for analysis.

Experimental Studies Of Multilayer Glass Structures On Compression, Compression With Bending And Simple Bending

2019 ◽  
pp. 22-30
Keyword(s):  
Cross Section ◽  
Cross Sections ◽  
Applied Load ◽  
Experimental Studies ◽  
Strength Properties ◽  
Research Topic ◽  
Normative Values ◽  
Laminated Glass ◽  
The Cross ◽  
Experimental Values

The work of multilayer glass structures for central and eccentric compression and bending are considered. The substantiation of the chosen research topic is made. The description and features of laminated glass for the structures investigated, their characteristics are presented. The analysis of the results obtained when testing for compression, compression with bending, simple bending of models of columns, beams, samples of laminated glass was made. Overview of the types and nature of destruction of the models are presented, diagrams of material operation are constructed, average values of the resistance of the cross-sections of samples are obtained, the table of destructive loads is generated. The need for development of a set of rules and guidelines for the design of glass structures, including laminated glass, for bearing elements, as well as standards for testing, rules for assessing the strength, stiffness, crack resistance and methods for determining the strength of control samples is emphasized. It is established that the strength properties of glass depend on the type of applied load and vary widely, and significantly lower than the corresponding normative values of the strength of heat-strengthened glass. The effect of the connecting polymeric material and manufacturing technology of laminated glass on the strength of the structure is also shown. The experimental values of the elastic modulus are different in different directions of the cross section and in the direction perpendicular to the glass layers are two times less than along the glass layers.

Cross Section Analysis of Cu Filled TSVs Based on High Throughput Plasma-FIB Milling

2012 ◽  
Author(s):  
Frank Altmann ◽  
Jens Beyersdorfer ◽  
Jan Schischka ◽  
Michael Krause ◽  
German Franz ◽  
...  
Keyword(s):  
High Resolution ◽  
Cross Section ◽  
High Throughput ◽  
Cross Sections ◽  
Electron Backscatter Diffraction ◽  
Grain Structure ◽  
Electron Backscatter ◽  
Section Surface ◽  
Backscatter Diffraction ◽  
Section Analysis

Abstract In this paper the new Vion™ Plasma-FIB system, developed by FEI, is evaluated for cross sectioning of Cu filled Through Silicon Via (TSV) interconnects. The aim of the study presented in this paper is to evaluate and optimise different Plasma-FIB (P-FIB) milling strategies in terms of performance and cross section surface quality. The sufficient preservation of microstructures within cross sections is crucial for subsequent Electron Backscatter Diffraction (EBSD) grain structure analyses and a high resolution interface characterisation by TEM.

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