bluuebunny/crossref_metadata_2025_split huggingface dataset:
3D biological imaging using TSRLM confocal microscopy
Authors: Loren Buhle E. , Himpens Bernard , Somlyo Avril V. , Shuman Henry , Somlyo Andrew P. 
Published: June 2020

Tandem scanning reflected light (TSRLM) confocal microscopy permits the imaging of optical sections through translucent material in real time. We are currently using a TSRLM microscope to examine the morphological components of biological tissue. Optical sections of either stained or unstained biological specimens can be rapidly obtained by a through focus series of a potentially dynamic specimen. Each optical section shown here is an average of 256 512X512x8 bit frames collected from a silicon intensified target camera at video rates.Unstained living frog muscle fibers were imaged with sufficient contrast to obtain 1μ optical sections traversing a total distance of approximately 80μ. Fig. 1 shows 4μ sections through a blood vessel. Frog red cells can be clearly seen in panels 1c-e. Frog muscle fibers were mounted and stretched to a defined length. The total fiber volume was easily imaged by progressive 1μ sections.
Cryosectioning plant roots prepared by high-pressure freezing
Authors: Crang R.E. , Mueller M. , Zierold K. 
Published: June 2020

Obtaining frozen-hydrated sections of plant tissues for electron microscopy and microanalysis has been considered difficult, if not impossible, due primarily to the considerable depth of effective freezing in the tissues which would be required. The greatest depth of vitreous freezing is generally considered to be only 15-20 μm in animal specimens. Plant cells are often much larger in diameter and, if several cells are required to be intact, ice crystal damage can be expected to be so severe as to prevent successful cryoultramicrotomy. The very nature of cell walls, intercellular air spaces, irregular topography, and large vacuoles often make it impractical to use immersion, metal-mirror, or jet freezing techniques for botanical material.However, it has been proposed that high-pressure freezing (HPF) may offer an alternative to the more conventional freezing techniques, inasmuch as non-cryoprotected specimens may be frozen in a vitreous, or near-vitreous state, to a radial depth of at least 0.5 mm.
Microprobe analysis of inclusion bodies related to two benzofuran derivatives
Authors: Benning T. L. , Ingram P. , Shelburne J. D. 
Published: June 2020

Two benzofuran derivatives, chlorpromazine and amiodarone, are known to produce inclusion bodies in human tissues. Prolonged high dose chlorpromazine therapy causes hyperpigmentation of the skin with electron-dense inclusion bodies present in dermal histiocytes and endothelial cells ultrastructurally. The nature of the deposits is not known although a drug-melanin complex has been hypothesized. Amiodarone may also cause cutaneous hyperpigmentation and lamellar lysosomal inclusion bodies have been demonstrated within the cells of multiple organ systems. These lamellar bodies are believed to be the product of an amiodarone-induced phospholipid storage disorder. We performed transmission electron microscopy (TEM) and energy dispersive x-ray microanalysis (EDXA) on tissue samples from patients treated with these drugs, attempting to detect the sulfur atom of chlorpromazine and the iodine atom of amiodarone within their respective inclusion bodies.A skin biopsy from a patient with hyperpigmentation due to prolonged chlorpromazine therapy was fixed in 4% glutaraldehyde and processed without osmium tetroxide or en bloc uranyl acetate for Epon embedding.
Scanning Electron Microscopy of the glomerulus in diabetic BB/S wistar rats
Authors: Kaufmann R. C. , Khosho F. K. , Amankwah K. S. 
Published: June 2020

Renal damage secondary to diabetes seems to be related to the severity and duration of the diabetes. In streptozotocin and alloxan-induced diabetic rats, renal disease is found only in those rats that have glycosuria and then only after the glycosuria has been present for many months. In these animals, the longer they have glycosuria, the more severe the renal damage. In our colony of BB/S Wistar rats, animals that are going. to become frankly diabetic demonstrate clinical diabetes before they begin spilling glucose in their urine. After glycosuria develops, the condition of the animals worsens; yet, the glucose tolerance tests(GTT) remain essentially unchanged. The purpose of this investigation was to study the animals' kidneys to discover if lesions are present at the onset of glycosuria and how severe the lesions are.Rats of our BB/S Wistar strain were used the day they developed glycosuria. Similarly aged non-diabetic animals were used as controls.
I. The morphology of interleukin-2 stimulated human peripheral blood mononuclear cells killing tumor Cellsin vitro
Authors: Hook G. R. , Greenwood M. A. , Barba D. , Ikejiri B. , Chen S. N. , Oldfield E. , Muul L. M. 
Published: June 2020

Culture of peripheral blood mononuclear cells (PBM) with recombinant interleukin-2 (rIL-2) results in lymphokine activated killer cells (LAK) that can kill fresh non-cultured, natural killer cell resistant tumor cells. Clinically, human LAK and rIL-2 immunotherapy can cause regression of some tumors in humans and preliminary trials using LAK cells to treat glioma patients are beginning . LAK effector cells are a subset of the LAK population that must physically contact target cells in order to kill them. In this and the following abstract, we report the morphology of LAK cells killing human glioma derived cellsin vitro.PBM cells were obtained from normal adult volunteers and stimulated in culture for 4 days with rIL-2 (Cetus, Emeryville, CA). Glioma-derived cells were obtained at surgery from a recurrent glioma patient and established as a cell line (SNB-92). SNB-92 cells were positive for glial fibrillary acidic protein and were used at passage 8.
II. The morphology of interleukin-2 stimulated human peripheral blood mononuclear cells killing tumor cells In Vitro
Authors: Greenwood M. A. , Hook G. R. , Barba D. , Ikejiri B. , Chen S. N. , Oldfield E. , Muul L. M. 
Published: June 2020

In the previous abstract, we presented data on the cytolysis of glioma-derived cells (SNB-92) by lymphokine activated mononuclear cells (LAK). In this abstract, we describe our transmission electron microscopy (TEM) study of the interaction of LAK cells and SNB-92 in vitro during a time sequence of 5 minutes to 4 hours. Co-cultures were fixed in situ by the direct addition to the culture medium of 2.5% glutaraldehyde and prepared in a routine manner for TEM.Within 30 minutes, some cells within the LAK population make contact with glioma cells (Fig 1). These LAK cells assume an elongated shape when they approach their target. The nucleus is asymmetrically located distal to the point of contact. The cellular organelles and microtubules orient towards the target cell. The plasma membranes of both cells bind closely together and form numerous interdigitations.
Ultrastructure of rat alveolar macrophages activated in situ by poly:IC
Authors: Klinkner A. M. , Weiss R. A. , Kelley A. , Bugelski P. J. 
Published: June 2020

Polyinosinic:polycytidylic acid is an inducer of interferon and a macrophage activator. We have found that intratracheal instillation of polyI:C (IT-pI:C) activates rat bronchoalveolar lavage cells (BAL) for a variety of functions. Examination of Giemsa stained, cytocentrifuge preparations showed that IT-pI:C induced a population of BAL not seen in resident BAL. The morphology of these cells suggested that they might be derived from blood monocytes. To test this hypothesis we have examined several populations of macrophages that had been stained for endogenous peroxidase activity as a marker of cells derived from the monocyte-macrophage lineage.Macrophages were obtained from Fischer 344 rats. Peritoneal exudate cells (PEC) were collected by lavage 4 days after i.p. injection of 20 ml 3% thioglycolate. Buffy coat monocytes were separated from venous blood from naive rats.
Immunogold labeling of protein kinase in muscle cells
Authors: Philpott D. E. , Hand A. , Mednieks M. , Kato K. , Stevenson J. 
Published: June 2020

Muscle loss during spaceflight remains a medical concern in prolonged space travel. We have studied changes in cyclic AMP-dependent protein kinase (ATP: protein phosphotransferase, EC 2.7.1.37; cAPK) levels in heart muscle after simulated weightlessness and after actual microgravity on Space Lab-3. Biochemical measurements were made in the soluble and the particulate fraction of disrupted muscle tissue, but did not identify the exact subcellular location of this enzyme. In order to establish localization of the protein kinase in the muscle cells of the rat, monoclonal antibodies to the regulatory (R) subunits of the type II isoenzyme were prepared and employed in a protein A-gold immunolabeling procedure.Both rat striated and heart muscle were fixed in 2.5% glutaraldehyde (GA), cacodylate buffer, for 1 hour. Similar samples were fixed in Triple Fix (T.F.) in cacodylate buffer for one week. Osmium was not used.
Immunoelectron microscopic localization of endoglucanase in the frozen thin sections of a hypercellulolytic mutant of Trichoderma reesei
Authors: Ganguly Tapan , Ghosh Bijan K. 
Published: June 2020

Endoglucanase (EG) is an enzyme of cellulase complex depolymerizing crystalline cellulose to cellulose microfibril fragments. T. reesei Rut-C30 is a catabolite repression resistant mutant which enhances endoplasmic reticulum (ER) and produces 15-20 times more endoglycanase than the wild type QM6a. About 90% of this enzyme is secreted.Purified secreted EG from three day old culture was used for polyclonal antibody production. The IgG was labeled with 15-18nm colloidal gold. EG secreting mycellia, grown in Vogel's medium with 1% Avicel PH101, were collected by centrifugation. This pellate was washed in sodium cacodylate buffer, fixed in tannic acid-glutaraldehyde mixture and cryoprotected with 30% glycerol. This material was placed on the specimen holders and frozen in liquid freon followed by liquid nitrogen. Frozen thin sections were prepared in a LKB Cryonova microtome using glass knives and collected on sucrose drops.
Vesicular system associated with spermiogenesis of bullfrog
Authors: Liu K. C. , Tsay S. F. 
Published: June 2020

In the histologic and electron microscopic study of the male reproductive system of bullfrog, Rana catesbeiana, a vesicular system associated with spermiogenesis was observed. It appeared in the lumenal space of the seminiferous tubule (Fig. 1), in the heads of spermatids (Fig. 2), associated with the chromatins of the spermatid (Fig. 4). As deduced from sections, this vesicular system consisted of vesicles of various size or a large group of waving and twisted tubules (Fig. 3), After routine procedure of treatment for electron microscopy, the lumens of both of the vesicles and tubules were electron lucent.In human, vesicles and vesicular system associated with reproductive cell and tissue were reported. In abnormal spermiogenesis, flower-like body, actually vesicles, and giant vesicle associated with the head of spermatid were observed. In both cases the number of vesicle was limited from a single one to a few.
Intracisternal microtubules in proximal tubule cells of duck kidney: A serendipitous finding
Authors: Bhatnagar M.K. , Snelgrove-Hobson S.M. , Prasada Rao P.V.V. 
Published: June 2020

Cytpplasmic microtubules, ubiquitous cell organelles, lie free in the cytosol without being compartmented off by membranes. In rare instances, however, intracisternal microtubules (ICM) have been reported in ganglionic neurons of aging dogs and in the proximal tubule cells (PTC) of rabbit kidneys. We report here the presence of ICM in the PTC of the kidneys of Peking ducks (Anas platyrhynhos).A total of 106 Peking ducks (24 males and 24 females of 9 months, 48 females of 15 months, 6 males of 30 months and 2 males and 2 females of 48 months of age) were anesthetized and exsanguinated. Pieces of kidney cortex were fixed in 4% glutaraldehyde in 0.5M phosphate buffer (pH 7.3) at 29°C for four hours. Samples were post fixed in 2% osmium tetroxide for two hours. One micron sections of Epon-embedded cortices were stained with toluidine blue and thin sections (70-70 nm) contrasted with uranyl acetate and lead citrate were examined with a JEOL 100-S electron microscope at 80 kV.
Deep etching of rapidly frozen cytoskeletons of motile fibroblasts
Authors: Heath Julian P. , Bard Frédérique 
Published: June 2020

The cytoskeleton of motile cultured cells is a complex three dimensional network of actin microfilaments, microtubules and intermediate filaments. We are using rapid freezing and deep etching to visualise this network in human and chick fibroblasts after detergent extraction. This method offers high resolution, high contrast images that retain three dimensionality with minimal artefact, and gives views of the cytoskeleton that differ from those obtained with more conventional techniques such as HVEM of thick plastic sections, or CTEM of critical point dried whole mounts.Human skin fibroblasts and chick embryo heart fibroblasts were grown on No. 1 glass coverslips and fixed and extracted with 0.1 M PIPES buffer pH 7 containing 0.25% glutaraldehyde, 0.5% Triton X-100, 1 mM MgCl2, 1 mM EGTA, 1 μg/ml phalloidin and 5 μg/ml taxol, for 30 s to 10 min at RT; further fixed in 2.5% glutaraldehyde in PIPES for 10 min, postfixed in 1% osmium in PIPES for 5 min and dehydrated in 70% ethanol.
Detection of viruses by electron microscopy: Increased sensitivity by direct micro-ultracentrifugation of samples
Authors: Trudel Alain R. , Trudel M. 
Published: June 2020

AirfugeR (Beckman) direct ultracentrifugation of viral samples on electron microscopy grids offers a rapid way to concentrate viral particles or subunits and facilitate their detection and study. Using the A-100 fixed angle rotor (30°) with a K factor of 19 at maximum speed (95 000 rpm), samples up to 240 μl can be prepared for electron microscopy observation in a few minutes: observation time is decreased and structural details are highlighted. Using latex spheres to calculate the increase in sensitivity compared to the inverted drop procedure, we obtained a 10 to 40 fold increase in sensitivity depending on the size of particles. This technique also permits quantification of viral particles in samples if an aliquot is mixed with latex spheres of known concentration.Direct ultracentrifugation for electron microscopy can be performed on laboratory samples such as gradient or column fractions, infected cell supernatant, or on clinical samples such as urine, tears, cephalo-rachidian liquid, etc..
Scanning Electron Microscopy of the vagina in diabetic BB/S Wistar rats
Authors: Khosho F. K. , Kaufmann R. C. , Amankwah K. S. 
Published: June 2020

The influence of diabetes on reproductive organs in humans and various other mammalian species, including the laboratory rat, is not well documented. We have previously shown that 13% of our BB/S Wistar rat colony demonstrated persistent estrous on vaginal smears. Spontaneously-diabetic BB Wistar rats of our colony demonstrate persistent diestrous on vaginal smears long before showing glucose in urine. In this report, we describe our findings on the ultrastructure of the vaginal epithelium of the frankly diabetic(FD) rat. The rats which started our colony were obtained from the Animal Resource Division of the Government of Canada.Scanning electron microscopy was used to examine specimens of vaginal surface epithelium with comparisons being made with specimens from nondiabetic controls. Cardiovascular perfusion with a buffered glutaraldehyde solution was used forin vivofixation of the cervix and vagina.
Modified gach technique vs critical point drying: Shrinkage analysis for SEM
Authors: Thompson B. , Sculov N. , Crang R.E. 
Published: June 2020

The use of co-polymerized glutaraldehyde-carbohydrazide (GACH) was proposed for specimen preparation in scanning electron microscopy (SEM) as a means of avoiding dehydration in organic solvents, and to provide dimensionally stable biological specimens through a process of air-drying. It has been assumed that shrinkage of specimens prepared by the GACH technique should be less than that of conventionally-prepared material by critical point drying (CPD). In a previous study, Bell has reported significant shrinkage of whole cells for SEM. This report compares cell shrinkage in GACH and CPD preparations.Fibroblasts from newborn rats were grown on collagen-coated glass cover-slips (with alpha numeric grids etched onto the surface of the coverslips) in Eagle's minimum essential medium + 10% fetal calf serum for 7 d. (3). Using an inverted microscope with phase-contrast optics, micrographs were taken of the cultures in their live state and 1 h. after fixation with 2.5% glutaraldehyde in Dulbecco's phosphate buffered saline (Figs. 1 and 3).
Relationship between various types of inclusions in the broncho-alveolar space of axolotl lung
Authors: Keyhani E. 
Published: June 2020

The surfactant system of the mexican axolotl Ambystoma mexicanum exhibited the same basic structures as those described for mammalian surfactant. Here we report subdivision of the surfactant inclusions, their interconversion and their frequency in the lung of the axolotl.1)Type I inclusion or tubular myelin represented 8% of the total inclusions. It consisted of square to rectangular grid patterns (Figs. 1, 4). Each square or rectangle shared its walls with four neighboring units. The lattice appeared to be composed of membranous elements of 6 nm, crossing in two directions perpendicular to each other. A centrally located dense line of 2 nm diameter with a fuzzy contour crossed the middle section of each rectangle in its long axis (Fig. 4). Each rectangle measured about 17 x 27 nm for internal perimeter and 27 x 36 nm for center to center spacing of its membrane element. The average size of type I inclusions was about 0.8 μm.
Backscattered electron signals of coated specimens at low and high detector take-off angles
Authors: Hejna Jan 
Published: June 2020

Backscattered electron (BSE) signals of differently coated specimens are studied at two ranges of BSE take-off angles. Specimens are coated with one layer of a high atomic number (high Z) material or with two layers: first of a high Z material and second of a low Z material. Experiments are carried out in a scanning electron microscope equipped with a tungsten filament.Fig. 1 shows an arrangement of detectors. A ring detector subtending angles from 0° to 20° in respect to the specimen surface and a disc detector subtending angles from 60° to 80° are used. Detectors contain plastic scintillators (102A, NE Technology). The first specimen consists of an Au layer on a Cu substrate and a part of it is covered with an additional Cu layer. Fig. 2 shows a dependence of BSE signals on the thickness of the Au layer and Fig. 3 on the thickness of the Cu layer.
Acoustic microscopy techniques for the inspection of integrated circuit devices and packages
Authors: Moore Thomas M. 
Published: June 2020

In the last decade, a variety of characterization techniques based on acoustic phenomena have come into widespread use. Characteristics of matter waves such as their ability to penetrate optically opaque solids and produce image contrast based on acoustic impedance differences have made these techniques attractive to semiconductor and integrated circuit (IC) packaging researchers.These techniques can be divided into two groups. The first group includes techniques primarily applied to IC package inspection which take advantage of the ability of ultrasound to penetrate deeply and nondestructively through optically opaque solids. C-mode Acoustic Microscopy (C-AM) is a recently developed hybrid technique which combines the narrow-band pulse-echo piezotransducers of conventional C-scan recording with the precision scanning and sophisticated signal analysis capabilities normally associated with the high frequency Scanning Acoustic Microscope (SAM). A single piezotransducer is scanned over the sample and both transmits acoustic pulses into the sample and receives acoustic echo signals from the sample.
Thin-section photomicrography
Authors: Holm Jack , Goss Rachel 
Published: June 2020

A fundamental problem in conventional light microscopy has long been the lack of image quality brought about by the use of sections which are thicker than the depth of field (DOF) of the microscope. These sections are used because “paraffin & steel” microtomy techniques preclude sections thinner than a few microns. The transmission electron microscope, however, requires section thicknesses of less than a micron. This requirement has resulted in the development of “epoxy & glass” microtomy techniques, and the ability to cut sections as thin as a few hundred Ångstroms. These thin sections are not routinely used for light microscopy because of the difficulty of preparation, the need for large section areas, and because of photomicrographic problems. The first two reasons may inhibit the use of thin sections in some situations, but the photomicrographic problems are surmountable.There is some disagreement in the literature regarding the actual DOF in photomicrographic situations.
New computer-controlled dual stage SEM—model DS-700
Authors: Ueda T. , Nakatsuka H. , Kimura T. , Buchanan R. 
Published: June 2020

The dual stage electron-optical column design of the DS-700 provides a unique combination of high resolution and flexible sample handling. The top stage accomodates a sample between the polepieces of a low aberration immersion objective lens, while the bottom stage accepts large samples below a conical second objective lens. This column configuration has been described elsewhere, and is therefore not the subject of this paper.A completely new digital electronic system with computer control of all operating parameters has been developed. A combined keyboard and menue control system has been adopted. Commonly used functions are assigned to a simple operating panel, with discrete functions which may be set and easily changed. Secondary functions are addressed using a menue display. In this manner, rapid simple operation is realized, while still providing extreme flexibility.A single high quality TV monitor displays real time and processed images. It is also used to display menues, various operating parameters, and vacuum system status.
The CM10-bioTWIN: a new CM microscope optimised for high contrast and analytical performance
Authors: de Jong Marc J.C. , Busing Wim M. , Otten Max T. 
Published: June 2020

In order to improve our understanding of life processes, morphological aspects of a specimen must be related to its chemistry. More and more morphological observations are combined with biochemistry, immunocytochemistry and analysis, demanding a different approach to specimen preparation than the one employed for morphological observation. In the case of immunocytochemistry the fixation is mild, while for X-ray analysis vapour fixation must be used to avoid loss of water-soluble substances. Heavy-element stains such as lead citrate or uranyl acetate normally cannot be used without introducing undesired and serious artefacts. The special specimen preparation procedures result in low TEM contrast, making it difficult to observe the morphology and hamper focussing especially if low beam intensities must be used as for beam-sensitive materials.Analytical TEMs typically are optimised for materials science applications, where low contrast is rarely a problem. These microscopes have low objective-lens aberrations and short focal lengths, giving low point resolution and nm-sized smallest spot sizes but at a expense of image contrast.