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At least 73 records · Page 4Linked to original sources
PubMed2002

Ribbons of semithin sections: an advanced method with a new type of diamond knife.

Complete series of semithin sections are imperative for 3-D reconstruction, but with traditional microtomy techniques it is difficult and time-consuming to trace stained and labeled structures. In the present study we introduce a method for making and collecting ribbons of semithin sections with a new, commercial available diamond knife (histo-jumbo-diamond knife, Diatome AG, Biel, Switzerland). The special feature of the diamond knife is the large water bath (boat) into which a glass slide can be dipped. The method has distinct advantages and the handling is simple. The resin block is trimmed into a truncated pyramid. Contact glue is applied to the leading face of the pyramid, which makes sections stick together to form a ribbon. Following sectioning, the ribbons are mounted onto glass slides and aligned in parallel. Stretching out and drying the ribbons on a hot plate is the final step of the method. Major advantages of this method are the perfect alignment of sections with identical orientation of structures, the completeness of series, and the significant saving of time. This facilitates tracing of stained and labeled structures, yielding quick 3-D reconstruction. Semithin sections can be cut from 0.5 to 2 micro m and several ribbons can be mounted side by side onto the slide. Two examples are presented to illustrate the advantages of the method.

Animals↗
PubMed1999

Combined ultramicrotomy for AFM and TEM using a novel sample holder.

A new sample holder that allows combined microtomy for atomic force microscopy (AFM) and transmission electron microscopy (TEM) is described. The main feature of this sample holder is a small central part holding the sample. This central part fits into the head of an atomic force microscope. AFM measurements can be performed with a sample mounted in this central part of the sample holder. This makes the alignment of a microtomed bulk sample unnecessary, and offers the opportunity of an easy and fast combined sample preparation for AFM and TEM.

Microscopy, Atomic Force↗
PubMed1955

Interpretation of electron micrographs of single and serial sections.

A method of securing serial sections for electron microscopy is described. Serial sections present certain anomalies of interpretation of a nature such that a complete and detailed three-dimensional reconstruction of the sectioned tissue cannot be made. These anomalies are discussed, as well as those which have been encountered in the interpretation of single sections. Observations of the following kinds have been made in an attempt to elucidate the interpretation of single and serial sections: differing methods of mounting adjacent sections, observation of the same section by high-angle stereoscopy, and examination of sections which have been shadowed prior to and subsequent to electron microscopy. It is found that the appearance of sections is independent of the choice of side to be placed against the formvar films. Stereoscopy shows that the appearance of fine structures is strongly dependent upon the direction of the penetrating electron beam with respect to the plane of the structures. Stereoscopy, combined with shadowing, shows quantitatively that extensive sublimation of polymer occurs upon normal exposure in the electron microscope. Observation of sections shadowed prior to electron microscopy indicates that varying amounts of material are removed between sections by the action of microtomy; i.e., it is probable that the sum of the thicknesses of several serial sections is considerably less than the total thickness of material removed from the block. It is believed that this effect, combined with the effect of sublimation, aids in explaining the failure of adjacent sections to exhibit continuity in their detailed structures.

Electrons↗
PubMed1960

Epoxy resins in electron microscopy.

A method of embedding biological specimens in araldite 502 (Ciba) has been developed for materials available in the United States. Araldite-embedded tissues are suitable for electron microscopy, but the cutting qualities of the resin necessitates more than routine attention during microtomy. The rather high viscosity of araldite 502 also seems to be an unnecessary handicap. The less viscous epoxy epon 812 (Shell) produces specimens with improved cutting qualities, and has several features-low shrinkage and absence of specimen damage during cure, minimal compression of sections, relative absence of electron beam-induced section damage, etc.-which recommends it as a routine embedding material. The hardness of the cured resin can be easily adjusted by several methods to suit the materials embedded in it. Several problems and advantages of working with sections of epoxy resins are also discussed.

Electrons↗
PubMed2006

Vitreous cryo-sectioning of cells facilitated by a micromanipulator.

Sectioning vitrified cells and tissues for cryo-electron microscopy is more challenging than room-temperature sectioning of plastic-embedded samples. As the sample must be kept very cold (<-130 degrees C) and because there is no liquid upon which the sections can float as they are cut, transferring the sections from the knife edge to a grid is one of the more difficult steps in the process. We employed a micromanipulator to hold and control the cryo-sections as they come off the knife. This allows slower cutting speeds than are typically used in vitreous cryo-sectioning and contributes to better control during cutting, which facilitates repeatable placement of a ribbon of sections onto a grid. The ribbon is kept under tension during the entire cutting process, which may decrease folding and/or compression, features that are inherent to vitreous sections. Furthermore, the added control afforded by this technique makes it easier for multiple ribbons to be placed on a single grid, thereby increasing the number of sections that can be examined and imaged during a microscopy session. It even allows for serial cryo-electron microscopy. As such, this approach is an advance in the cryo-microtomy of vitreous sections.

Cryoelectron Microscopy↗
PubMed2006

Prevalence of different types of accessory canals in the furcation area of third molars.

BACKGROUND: Although there are several studies that show the prevalence and diameter of accessory root canals in the furcation area, there is a scarceness of studies that observe the trajectory and different types of cavo-interradicular canals. The aim of this study was to verify the prevalence of the different morphologic types of accessory canals in the furcation region in an attempt to show their trajectories. METHODS: Forty submerged mandibular third molars were used, which were extracted and decalcified so that the microtomy procedure in the mesio-distal axial plane could be performed, obtaining semiserial sections with thicknesses of 5 microm. The sections were stained with hematoxylin and eosin and observed under optical microscopy. RESULTS: All of the morphologic types were found, whereas the proper accessory canals, type-A canals, were present in 10% of the specimens. The most prevalent canals were the sealed ones (type D), with a prevalence of 87.5%, followed by the blind ones (type B), with a prevalence of 75%. The loop accessory canals (type C), observed in only 5% of the teeth, were the least prevalent ones. CONCLUSIONS: Different morphologic types of accessory canals were found in the furcation area of submerged mandibular molars. The histologic method was effective to show the canal trajectories.

Coloring Agents↗
PubMedDate not supplied

Cryoultramicrotomy as a preparative method for x-ray microanalysis in pathology.

The role of ion shifts in cell injury has become one of the most intriguing fields of application of X-ray microanalysis in pathology. In principle, cryo(ultra)-microtomy is the best preparative method for X-ray microanalysis of diffusible substances. In this review, the following points are discussed: the choice of freezing method and coolant, the use of high molecular weight polymer cryoprotectants and their possible physiological effects the choice of the cryosectioning temperature, and techniques for handling and transfer of sections. Experiences with the sectioning of cryoprotected tissue are presented. Finally, other preparative techniques for microanalysis are compared to cryoultramicrotomy.

Animals↗
PubMed1989

[Heart growth in the human embryonic period (a study related to the 2d month of gestation)].

The growth of the heart was studied in 27 human staged embryos of post-somitic period (stages from 15 to 23, Carnegie Institution of Washington). The volume of the heart was determined multiplying the cardiac profiles areas (evaluated by point counting planimetry on serial sections) by the thickness of microtomy. Cardiac volume increase was examined relative to age, to crownrump length, and to body weight. The data were transformed into logarithms and bivariate relations were analysed according allometric equations of the form (log y = k log x + log b). The data were fitted according to three techniques: least squares, major axis, and reduced major axis. The slopes of the allometric regressions indicate positive allometry of the cardiac volume relative to embryonic age and to C-R length, and isometry relative to body weight.

Cardiac Volume↗
PubMed1986

The diamond knife "semi": a substitute for glass or conventional diamond knives in the ultramicrotomy of thin and semi-thin sections.

The diamond knife "semi" for ultramicrotomes was originally designed by its manufacturer (DIATOME S.A.), for cutting semi-thin sections from 0.2 micron to 2.0 micron. Cutting tests of Epon-embedded material (nervous system, myelin sheat) with this knife have shown that the quality of semi-thin sections is equivalent or better than that obtained with a glass knife, and much time could be saved during the microtomy of serial sections. The quality of thin sections (0.07 micron to 0.12 micron) is excellent and comparable to that obtained with a conventional diamond knife. Furthermore, when adjacent sections are cut thin and semi-thin (for immunochemistry or high voltage microscopy), both are excellent in that they are of uniform thickness. In conclusion, this tool has advantages for both light microscopy and transmission electron microscopy.

Animals↗
PubMedDate not supplied

[A unified series of microtomes for biology and medicine on the basis of rotational microtome].

A block-module constructor for the assembly of rotation microtomes is described. The microtomes are used for cutting fixed and native tissues. The basic rotation microtome is the main element of the microtome constructor. A set of changeable modules enables one to assemble microtomes on the basis of the basic rotation microtome for cutting fixed embedded tissue, 5-20 microns thick, fixed nonembedded tissue, 75-300 microns thick, native tissue, 150-600 microns thick, as well as a microtome-vibratome (the thickness of sections is 50-200 microns). A universal microtome combining all the mentioned-microtomes may be designed. The problems of fixed and native tissue microtomy are discussed in the paper.

Biology↗
PubMed1997

Improved methods for making and using glass knives.

We have observed over time that the right side of a glass knife is the optimal cutting edge for microtomy if the counterpiece (heel opposite the edge) is controlled within 1 mm. The right cutting edge has been considered the "saw toothed" side and has not been used for ultrathin sectioning. We have observed that the right cutting edge is sharper and more durable than the left. Light and scanning electron microscopy were used to observe the cutting edge, and transmission electron microscopy was used to examine semithin and ultrathin sections of animal and plant tissues cut by the right and left sides of the cutting edge. The results indicate that the cutting edge becomes sharper and more durable from left to right. Both the quality and efficiency of ultrathin sectioning is improved by using the right cutting edge.

Animals↗
PubMed1997

An assessment of the distortion of arteries due to sectioning in endomyocardial biopsies.

Arteries are usually studied morphometrically after pressurized fixation and resin embedding. These procedures are impracticable when dealing with diagnostic biopsies. The accuracy of arterial morphometry is determined partly by the degree of tissue distortion during section preparation. The axial ratios of 7340 arteries were measured in 353 endomyocardial biopsies from 23 patients and then compared with those expected from mathematical modelling. An excess of elliptical arteries was found. The distribution of orientation of the long axes of these best fitted a simulated 10 per cent linear distortion in the direction of microtomy. In conclusion, these results suggest that although there is some tissue distortion during sectioning, useful data may be obtained from morphometry of arteries in routinely processed endomyocardial biopsies.

Biopsy↗
PubMed1992

Preparation of individual electrically and video-recorded eggs for integrated temporal and electron microscopic analyses.

A method for correlative studies of early fertilization events that integrates techniques of intracellular electrophysiological recording, video-imaging, and electron microscopy is described. A key feature of the method is its ability to identify the fertilizing sperm and to record the moment of egg excitation. Since the site of gamete interaction is recognizable throughout all stages of preparation, difficulties associated with locating the site of fertilization and determining specimen orientation for microtomy and electron microscopic examination are eliminated. Virtually all samples yield useful information. An example of interacting gametes fixed 4 sec after initiation of the fertilization potential and serial sectioned is described. The method is applicable to systems other than fertilizing eggs when functional, temporal, and spatial relationships of individual cells need to be correlated with changes in ultrastructure.

Animals↗
PubMed2007

Novel procedure for high-fidelity tendon histology.

A novel method to obtain high-quality histologic sections of rat tendon for light microscopic study is presented. This approach utilizes non-deleterious dehydrating and clearing solvents and a hydrophobic acrylic (methyl methacrylate) resin. This methodology avoids processing and microtomy artifacts common with routine paraffin wax techniques and overcomes specimen size limitations associated with hydrophilic (e.g., glycol methacrylate) resin histologic methods. These novel histologic processing techniques facilitate the reliable assessment of tendons' cellular and matrix components and can be readily adapted to morphologic studies of damage, healing, and repair.

Acrylic Resins↗
PubMedDate not supplied

Cryo-planing for cryo-scanning electron microscopy.

In the past decade, investigators of cryo-planing for low-temperature scanning electron microscopy (cryo-SEM) have developed techniques that enable observations of flat sample surfaces. This study reviews these sample preparation techniques, compares and contrasts their results, and introduces modifications that improve results from cryo-planing. A prerequisite for all successful cryo-planing required a stable attachment of the specimen to a holder. In most cases, clamping with a screw mechanism and using indium as space-filler sufficed. Once this problem was solved, any of three existing cryo-planing methods could be used to provide successful results: cryo-milling, microtomy in a cold room, and cryo-ultramicrotomy. This study introduces modifications to the cryo-planing technique that produces flat surfaces of any desired plane through a specimen. These flat surfaces of frozen, fully hydrated samples can be used to improve observations from cryo-SEM as well as to enhance results from x-ray microanalysis and (digital) image analysis. Cryo-planing results of chrysanthemum (Dendranthema x grandiflorum Tzvelev) stems, hazel (Corylus avelane L.) stems, and repeseed (Brassica napus L.) pistils are presented to illustrate the use of the planing method on fibrous, hard, and delicate materials, respectively.

Cryoelectron Microscopy↗
PubMed1984

Structural studies of glycerinated skeletal muscle. I. A-band length and cross-bridge period in ATP-contracted fibers.

An electron microscope study is reported of structural changes during ATP-induced contraction of glycerinated rabbit psoas. In the absence of ATP, A-band length is constant at sarcomere lengths above 1.9 micron, with average length of 1.54 mu. In ATP-treated fibers, A-band length is also constant at sarcomere lengths above 2.0 microns, but the apparent length of A-band decreases to approximately 1.3 micron, as sarcomere length decreases from 1.9 micron to 1.5 mu. The occurrence of short A-bands cannot be attributed to crumpling of thick filaments against Z-lines, since I-bands remain patent; nor to the presence of heterogeneous filaments, since resting muscle does not show comparable heterogeneity, nor to compressive artifacts, which are minor when knife edge is oriented parallel with fiber axis during microtomy . The decrease of A-band length appears related, at least in part, to disarray of terminal cross-bridges as the thick filaments encroach upon the N-line, a structure which becomes evident within the I-band during contraction of glycerinated fibers. In preliminary studies, optical transforms of A-bands from individual sarcomeres reveal a characteristic myosin layer-line pattern as low as 1.5 micron sarcomere length. A cross-bridge repeat of 143 A is obtained for sarcomeres above 1.6 micron length; however, an appreciable proportion of sarcomeres in the range from 1.5 micron to 1.9 mu length generate meridional reflections less than 143 A, and as low as 130 A.

Adenosine Triphosphate↗
PubMed1990

A numerical procedure for choosing effective, low toxicity plasticizers for glycol methacrylate embedding.

A numerical method for selecting low toxicity plasticizers for glycol methacrylate (GMA) embedding mixtures is described. It involves use of various numerical parameters, namely; molecular weight, melting point, solubility parameter, and toxicity data. On the basis of these parameters, nine plasticizers were selected. Their effects on microtomy and tissue processing, and also their influence on tissue morphology and staining, were investigated. For tissue processed at low temperatures into GMA, using ethanol dehydration, the following compounds were as satisfactory as 2-butoxyethanol whilst being less toxic: 2-isopropoxyethanol, 2-(2-methoxyethoxy)ethanol, 2-(2-ethoxyethoxy)ethanol, 2-(n-butoxyethoxy)ethanol. However for tissues processed using the plasticizer as dehydrating agent, the optimum plasticizers were 2-isopropoxyethanol, 2-(2-methoxyethoxy)ethanol and 2-(2-ethoxyethoxy)ethanol. It is possible to give a numerical specification of the preferred plasticizers, and for the first procedure this is: a solubility parameter in the range 21-26 J0.5 cm-1.5 or higher, a melting point well below 0 degree C, and a rat oral LD50 greater than 12.52 mmol kg-1. It was also possible to analyse the differential effects of the dehydrating agents on histochemical and enzyme histochemical staining on a numerical basis, using a structure-activity relations approach.

Animals↗
PubMed1976

Changes in alkaline phosphatase activity in periosteal cells in healing fractures.

The quantitative changes in alkaline phosphatase activity in the periosteal cells close to the fracture in rat metatarsal bones has been measured during the first 5 days postfracture. This study has been made possible by two technological advances, firstly the development of cryostat microtomy for cutting unfixed, undemineralised bone, and secondly the use of scanning and integrating microdensitometry for quantifying the activity in each periosteal cell. The results showed a loss of alkaline phosphatase activity close to the fracture site, with activity rising to normal values 0.8-1.0 mm from the site. No alkaline phosphatase activity was found in the cells which proliferated from the periosteum. It is suggested that reduced glutathione could cause such inhibition.

Alkaline Phosphatase↗