Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Microscopy, Video”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 757 records · Page 42Linked to original sources
PubMed2004

Distribution of single DNA molecule electrophoretic mobilities in semidilute and dilute hydroxyethylcellulose solutions.

The distribution of center of mass electrophoretic mobility mobilities and normalized migration time of up to 1080 lambda DNA molecules per experiment were measured in both semidilute hydroxyethylcellulose HEC/0.5 x Tris-borate-EDTA (TBE) solutions and dilute HEC/0.5 x TBE solution by high-speed video microscopy. Measurements were made microscopically over a short migration distance in homogeneous DNA HEC/0.5 x TBE solution and after electrophoretic migration of a plug of DNA through 7 cm. Video at 120 frames/s (semidilute HEC solution) and 236 frames/s (dilute HEC solution) allowed visualization with adequate resolution for single molecule mobility measurements. The electrophoretic migration times and band shapes predicted from the measurements corresponded well with those measured by conventional capillary electrophoresis (CE) in both semidilute and dilute HEC. In semidilute solution, the band width predicted by a square root of time scaling is in good agreement with the results of conventional CE. However, in dilute solution the precision of the measurements was not good enough to allow scaled estimates of band widths.

Cellulose↗
PubMed1991

Quantitative fluorescence techniques for the determination of local microtubule polymerization equilibria in cultured neurons.

The local control of intracellular microtubule polymerization equilibria has been hypothesized to be an important factor in the determination of neurite extension and other examples of cellular asymmetry. Provided that the quantum yield of the fluorophore remains constant, the combination of fluorescent analogue cytochemistry, differential extraction protocols, and quantitative video microscopy makes it possible to measure local fractions of cytoskeletal protein in polymer, even when it is impossible to resolve individual fibrils of the polymer. We have developed appropriate quantitative video microscopic techniques for measuring the fluorescence of a fluorescent analogue-injected neurite before and after extraction under microtubule-stabilizing conditions. We have used these methods to demonstrate that tetramethylrhodamine-n-hydroxysuccinimide tubulin is an appropriate fluorescent analogue, allowing us to measure fractions of tubulin in polymer locally within PC12 neurites. As would be expected, the fraction of tubulin fluorescent analogue in polymer approaches 1.0 in neurites exposed to the microtubule-stabilizing drug taxol and is close to 0 in neurites injected and extracted in the cold, or extracted under microtubule-destabilizing conditions. We have, therefore, developed a tool that allows us to measure microtubule polymerization equilibria out the neurites of cells in culture, which will allow us to test hypotheses that factors which affect neurite outgrowth do so by means of effects on microtubule polymerization equilibria.

Adrenal Gland Neoplasms↗
PubMed1990

Ophidian kidney preparation for the measurement of glomerular dynamics in real-time.

A new ophidian kidney preparation is described which allows the measurement of glomerular arteriolar diameters, glomerular blood flow, and glomerular capillary pressure in real time. Intravital epifluorescence video microscopy is utilized to observe and record blood flow in the renal microcirculation in the garter snake, Thamnophis sirtalis. Carotid, renal artery and glomerular capillary pressures are recorded digitally on the video recording simultaneously with the images with an eight-channel video data recorder, maintaining synchrony of all data. On the video replay, glomerular arteriolar diameters are measured with a video micrometer and RBC velocity determined with the video dual-slit method. Blood flow is continuously calculated from the diameter and RBC velocity measurements. The continuous and simultaneous measurements of the pressure gradient across the afferent limb of the glomerular circulation, the renal artery to glomerular capillary pressure drop, and the rate of glomerular blood flow allow the continuous calculation of afferent glomerular arteriolar resistance in real time. This is the first demonstration of these capabilities in a vertebrate kidney.

Animals↗
PubMed2001

Comparative study on motility of the cultured fetal and neonatal dermal fibroblasts in extracellular matrix.

One of the differences between fetal and adult skin healing is the ability of fetal wounds heal without contraction and scar formation. Extracellular matrix (ECM) provides a substratum for cells adhesion, migration, and proliferation and can directly influence the form and function of cells. As motility is essential for many important biological events, including wound healing, inflammatory response, embryonic development, and tumor metastasis, this study was designed to compare the motilities cultured dermal fetal and neonatal fibroblasts in the extracellular matrix. The motility of cultured fetal and neonatal fibroblasts was compared using a video-microscopy system that was developed in combination with a self-designed CO2 mini-incubator. To determine migration speed, cells were viewed with a 4X phase-contrast lens and video recorded. Images were captured using a color CCD camera and saved in 8-bit full-color mode. We found that cultured fetal fibroblasts move faster than neonatal fibroblast on type I collagen (fetal fibroblast, 15.1 micrometer/hr; neonatal fibroblast, 13.7 micrometer/hr), and in fibronectin (fetal fibroblast, 13.2 micrometer/hr; neonatal fibroblast, 13.0 micrometer/hr) and hyaluronic acid (fetal fibroblast, 11 micrometer/hr; neonatal fibroblast, 9.8 micrometer/hr).

Cell Movement↗
PubMed1987

Influence of the calcium ionophore A23187 on rat egg behavior and cortical F-actin.

Rat eggs treated with the calcium ionophore A23187 and subjected to long-term observation by phase microscopy were found to undergo many developmental changes that are normally associated with fertilization. These included cortical granule exocytosis and the abstriction of the second polar body. In addition, time-lapse video microscopy revealed that, unlike untreated eggs, whose surfaces remained relatively immotile, the ionophore-treated eggs underwent a lengthy period of surface undulatory activity. Since all of these events were remarkably similar in timing and morphology to those seen in fertilized eggs, we conclude that A23187 is capable of activating rat eggs. Using NBD-phallacidin, the distribution of F-actin in ionophore-activated eggs was determined. During most of the postactivation period the eggs possessed an uninterrupted, uniform band of polymerized actin encompassing the entire cortex of the egg. However, during a discrete 1.5-h period after the formation of the second polar body, an area adjacent to the region of polar body abstriction exhibited more intense staining than the rest of the cortex. Cytochalasin B treatment caused a dramatic reduction and/or rearrangement in cortical NBD-phallacidin staining in activated eggs as compared to activated controls not exposed to the drug. We observed that all the developmental changes described above could be produced in the absence of exogenous calcium, suggesting that the rat egg possesses internal stores of calcium sufficient to elicit an activational response. We conclude that the ionophore-induced release of free calcium ions into the cytosol stimulates many of the developmental changes that are normally seen during fertilization. These results indicate that calcium influx and cytoskeletal activity are correlated during the activation of this animal egg.

Actins↗
PubMed2006

Exploring bovine pancreatic trypsin inhibitor phase transitions.

This paper presents an investigation of the phase diagram of BPTI (bovine pancreatic trypsin inhibitor)/350 mM KSCN at pH 4.9 by direct observation and numerical simulations. We report optical microscopy and light and X-ray scattering experiments coupled with theoretical data analysis using numerical tools. The phase diagram is thoroughly determined, as a function of temperature. Two polymorphs are observed by video microscopy and their solubility measured. In this phase diagram, the liquid-liquid phase separation (LLPS) is metastable with respect to the solid-liquid phase separation. Above the T(L-L) boundary curve, solutions are composed of a mixture of BPTI monomers and decamers. Attractive interactions are stronger between decamers than between monomers. Below the T(L-L) boundary curve, the dense phase is highly concentrated in protein and composed of BPTI decamers alone. Thus, the driving force for liquid-liquid or liquid-solid phase separation is the attraction between decamers at low pH. The structure factors of the dense phases are characteristic of repulsive dense phases because of a hard sphere repulsion core, meaning that in the dense phase proteins are actually in contact (interparticle distance of 53 A). In agreement with the Oswald rule of stages, LLPS occurs prior to and impedes the solid nucleation.

Animals↗
PubMed2001

Direct, high-resolution measurement of furrow stiffening during division of adherent cells.

It is unclear whether cell division is driven by cortical relaxation outside the equatorial region or cortical contractility within the developing furrow alone. To approach this question, a technique is required that can monitor spatially-resolved changes in cortical stiffness with good time resolution. We employed atomic force microscopy (AFM), in force-mapping mode, to track dynamic changes in the stiffness of the cortex of adherent cultured cells along a single scan-line during M phase, from metaphase to cytokinesis. Video microscopy, which we used to correlate the AFM data with mitotic events identified by light microscopy, indicated that the AFM force-mapping technique does not perturb dividing cells. Here we show that cortical stiffening occurs over the equatorial region about 160 seconds before any furrow appears, and that this stiffening markedly increases as the furrow starts. By contrast, polar relaxation of cells does not seem to be an obligatory event for cell division to occur.

Animals↗
PubMed1990

Measurements of blood flow to individual glomeruli in the ophidian kidney.

Continuous measurements of the instantaneous rate of blood flow to individual glomeruli in a normal vertebrate kidney were made in the garter snake Thamnophis sirtalis. Epifluorescence video microscopy was used to visualize and record blood flow in the afferent arterioles of superficial nephrons. The dual-slit method was used for the determination of red blood cell (RBC) velocity from the video replay. Simultaneous measurements of the vessel diameter allowed the continuous determination of the instantaneous rate of blood flow. A total of 100 glomeruli was surveyed in 12 animals. These glomeruli displayed both constant and highly variable rates of blood flow, with 21% of all nephrons displaying intermittent glomerular perfusion. The mean single-nephron blood flow rate (SNBFR) for all individuals was 23.9 +/- 10.3 (SD) nl/min (n = 12). The percentage of nephrons with intermittent flow for an individual animal increased significantly with increasing plasma osmolality. Intermittency was associated with low SNBFR values; SNBFR averaged 13.5 +/- 10.2 (SD) nl/min (n = 21) in intermittent nephrons and 29.2 +/- 19.0 (SD) nl/min (n = 79) in continuous flow nephrons, the difference being significant (P less than 0.001). Nephrons with continuous perfusion displayed a much greater range of SNBFR values than intermittent nephrons. This suggests that, although changes in whole kidney glomerular filtration rate (GFR) in reptiles need not involve glomerular intermittency, intermittency may lower GFR.

Animals↗
PubMed1997

Characterization of influenza virus-induced death of J774.1 macrophages.

The mechanism and role of influenza virus (IV)-induced pathogenesis of macrophages during respiratory infection are ill defined. Reported here are findings on IV-induced cytopathic effects (CPEs) for an in vitro experimental system using the murine macrophage cell line J774.1. CPE was elicited by 0.2 or greater multiplicity of infection (m.o.i.). CPEs showed a lag of 6-8 h postinfection and occurred most rapidly between 6 and 12 h. J774.1 cells did not support productive IV replication, but immunofluorescence demonstrated that IV protein synthesis occurred. Light microscopy and DNA staining showed that after death cells had very condensed cytoplasm and nuclei. Cell remnants were surrounded by intact plasma membrane (PM) as demonstrated by exclusion of a membrane-impermeant dye. Time-lapse video microscopy recordings between 6 and 10 h postinfection showed sequential structural changes, including previously undescribed events. Notable changes were a rapid cytokinesis (zeiosis; "cell boiling"), followed by nuclear shrinkage, and an unusual transient blebbing of the PM. DNA fragmentation occurred after 12 h, producing a wide size range. UV-inactivated virus failed to induce CPEs, and CPE was blocked by amantadine. N-Acetylcysteine and pyrrolidine dithiocarbamate, but not other inhibitors of reactive oxygen intermediates, reduced or blocked the CPE. Most changes observed are those attributed to apoptotic processes rather than necrotic cell death. The kinetics and inhibitor effects suggest that IV infection and replication must be initiated to activate CPEs.

Acetylcysteine↗
PubMed1996

Synchronous measurements of blood pressure and red blood cell velocity in capillaries of human skin.

We have devised a system for analyzing the temporal coherence of capillary pressure (CP) and capillary red blood cell velocity (CBV) variations in humans. The system is designed to measure human blood pressure in skin capillaries by direct cannulation while simultaneously measuring red blood cell velocity in the same capillary by video microscopy. The servo-nulling pressure measurement system allows the dynamic recording of capillary pressure with a flat frequency response of about 12 Hz. Computerized data acquisition is synchronized with the frame code of a U-matic video recorder, which records the capillaroscopic picture for later computerized off-line analysis of capillary red blood cell velocity. Measurements of simultaneously recorded CP and CBV in healthy volunteers show synchronized pulsation. Minimal pulsatile variations in CP values result in marked variations of CBV. In addition, minor fluctuations (3-4 cycles per minute) in CP are accompanied by marked changes in CBV. This system may help provide information about microvascular pathophysiology in skin diseases before and after treatment.

Adult↗
PubMed1989

Telepathology. Long-distance diagnosis.

Telepathology is defined as the practice of pathology at a distance, by visualizing an image on a video monitor rather than viewing a specimen directly through a microscope. Components of a telepathology system include the following: (1) a workstation equipped with a high-resolution video camera attached to a remote-controlled light microscope; (2) a pathologist workstation incorporating controls for manipulating the robotic microscope as well as a high-resolution video monitor; and (3) a telecommunications link. Progress has been made in designing and constructing telepathology workstations and fully motorized, computer-controlled light microscopes suitable for telepathology. In addition, components such as video signal digital encoders and decoders that produce remarkably stable, high-color fidelity, and high-resolution images have been incorporated into the workstations. Resolution requirements for the video microscopy component of telepathology have been formally examined in receiver operator characteristic (ROC) curve analyses. Test-of-concept demonstrations have been completed with the use of geostationary satellites as the broadband communication linkages for 750-line resolution video. Potential benefits of telepathology include providing a means of conveniently delivering pathology services in real-time to remote sites or underserviced areas, time-sharing of pathologists' services by multiple institutions, and increasing accessibility to specialty pathologists.

Computer Communication Networks↗
PubMedDate not supplied

In vivo and in vitro analysis of the effectiveness of doxorubicin combined with temporary arterial occlusion in liver tumors.

PURPOSE: The authors evaluated the effects of daunomycin (daunorubicin)--an analogue of doxorubicin--ethiodized oil, and arterial occlusion on an in vitro hepatoma analogue and on in vivo rat liver tumors. MATERIALS AND METHODS: A human Sk hepatoma cell monolayer sandwich system was used to determine uptake of 3H-daunomycin under normoxic/hypoxic conditions with use of autoradiography. Fluorescence microscopy was used to evaluate the biodistribution of doxorubicin in cell cultures (human Sk hepatoma and colon carcinoma). Microvascular flow adjacent to and within liver tumors and the intrahepatic effects of doxorubicin and ethiodized oil were studied with in vivo video microscopy on exteriorized rat livers containing peripheral hepatomas. RESULTS: Increased uptake of 3H-daunomycin by hepatoma cells occurred under hypoxic conditions. Intrahepatic arterial administration of ethiodized oil caused temporary occlusion of peripheral sinusoids following passage through arterioportal anastomoses. Tumors received portal venous and neovascular blood supply and ethiodized oil occluded but did not enter the narrow neovasculature perfusing the tumors. CONCLUSION: Hypoxia increases uptake of 3H-daunomycin by human Sk hepatoma and colon carcinoma cell cultures. Selective hepatic arterial occlusion (and perhaps the resultant hypoxia) may facilitate increased uptake of doxorubicin analogues into liver tumors. Hepatomas receive both arterial and portal venous blood supply, and ethiodized oil reaches the tumor via arterioportal anastomoses that perfuse the tumor periphery.

Animals↗
PubMed1998

Time-lapse microscopy reveals unique roles for kinesins during anaphase in budding yeast.

The mitotic spindle is a complex and dynamic structure. Genetic analysis in budding yeast has identified two sets of kinesin-like motors, Cin8p and Kip1p, and Kar3p and Kip3p, that have overlapping functions in mitosis. We have studied the role of three of these motors by video microscopy of motor mutants whose microtubules and centromeres were marked with green fluorescent protein. Despite their functional overlap, each motor mutant has a specific defect in mitosis: cin8Delta mutants lack the rapid phase of anaphase B, kip1Delta mutants show defects in the slow phase of anaphase B, and kip3Delta mutants prolong the duration of anaphase to the point at which the spindle becomes longer than the cell. The kip3Delta and kip1Delta mutants affect the duration of anaphase, but cin8Delta does not.

Anaphase↗
PubMed1984

Two forms of cerebellar glial cells interact differently with neurons in vitro.

Specific interactions between neurons and glia dissociated from early postnatal mouse cerebellar tissue were studied in vitro by indirect immunocytochemical staining with antisera raised against purified glial filament protein, galactocerebroside, and the NILE glycoprotein. Two forms of cells were stained with antisera raised against purified glial filament protein. The first, characterized by a cell body 9 microns diam and processes 130-150 microns long, usually had two to three neurons associated with them and resembled Bergmann glia. The second had a slightly larger cell body with markedly shorter arms among which were nestled several dozen neuronal cells, and resembled astrocytes of the granular layer. Staining with monoclonal antisera raised against purified galactocerebroside revealed the presence of immature oligodendroglia in the cultures. These glial cells constituted approximately 2% of the total cell population in the cultures and, in contrast to astroglia, did not form specific contacts with neurons. Staining with two neuronal markers, antisera raised against purified NILE glycoprotein and tetanus toxin, revealed that most cells associated with presumed astroglia were small neurons (5-8 microns). After 1-2 d in culture, some stained neurons had very fine, short processes. Nearly all of the processes greater than 10-20 micron long were glial in origin. Electron microscopy also demonstrated the presence of two forms of astroglia in the cultures, each with a different organizing influence on cerebellar neurons. Most neurons associated with astroglia were granule neurons, although a few larger neurons sometimes associated with them. Time-lapse video microscopy revealed extensive cell migration (approximately 10 microns/h) along the arms of Bergmann-like astroglia. In contrast, cells did not migrate along the arms of astrocyte-like astroglia, but remained stationary at or near branch points. Growth cone activity, pulsating movements of cell perikarya, and ruffling of the membranes of glial and neuronal processes were also seen.

Animals↗
PubMed1996

Plant mitosis promoting factor disassembles the microtubule preprophase band and accelerates prophase progression in Tradescantia.

The regulation of mitosis in higher plant cells has been investigated by microinjecting protein kinase from the metaphase-arresting (met1) mutant of Chlamydomonas. Biochemical characterization of this enzyme complex confirms the presence of a p34cdc2/cyclin B-like kinase. The enzyme was injected into living stamen hair cells of Tradescantia virginiana in which microtubules (MTs) were visualized using fluorescent analogue cytochemistry and confocal laser scanning microscopy. Microinjection of this p34cdc2/cyclin B-like kinase caused rapid disassembly of the preprophase band of MTs but not of interphase-cortical, spindle or phragmoplast MTs. Effects of the enzyme on the cytomorphology of live prophase cells were also monitored using video microscopy. We found that injection of this enzyme accelerated chromatin condensation and nuclear envelope breakdown. This indicates the presence and function in plants of an enzyme that can initiate nuclear division similar to the maturation or mitosis promoting factor (MPF) of animal cells. These studies provide the first direct evidence that the mitotically-active form of plant MPF can drive disassembly of preprophase band MTs, chromosome condensation and initiation of mitosis in plant cells.

Animals↗
PubMed2004

Evaluation of rapid volume changes of substrate-adherent cells by conventional microscopy 3D imaging.

Precise measurement of rapid volume changes of substrate-adherent cells is essential to understand many aspects of cell physiology, yet techniques to evaluate volume changes with sufficient precision and high temporal resolution are limited. Here, we describe a novel imaging method that surveys the rapid morphology modifications of living, substrate-adherent cells based on phase-contrast, digital video microscopy. Cells grown on a glass substrate are mounted in a custom-designed, side-viewing chamber and subjected to hypotonic swelling. Side-view images of the rapidly swelling cell, and at the end of the assay, an image of the same cell viewed from a perpendicular direction through the substrate, are acquired. Based on these images, off-line reconstruction of 3D cell morphology is performed, which precisely measures cell volume, height and surface at different points during cell volume changes. Volume evaluations are comparable to those obtained by confocal laser scanning microscopy (DeltaVolume < or = 14%), but our method has superior temporal resolution limited only by the time of single-image acquisition, typically approximately 100 ms. The advantages of using standard phase-contrast microscopy without the need for cell staining or intense illumination to monitor cell volume make this system a promising new tool to investigate the fundamentals of cell volume physiology.

Cell Adhesion↗
PubMed1981

[New ways of studying the cutaneous microcirculation in clinical medicine (author's transl)].

Transcutaneous video microscopy opens a way to measure capillary red blood cell speed, to analyzed transcapillary diffusion and pericapillary distribution of Na-fluorescein injected i.v. and to depict the superficial lymphatic capillaries by subepidermal microinjection of FITC-labelled dextran in human skin. The dynamic phenomena may be quantitiated by video densitometers provided the information is stored on video tape. Already, some physiological and clinical data have been obtained. The velocity pattern of erythrocytes in nailfold capillaries is continuous or intermittent with flow stops. Standardized cold provocation tests are useful to evaluate Raynaud's disease. In patients with scleroderma the transcapillary passage of Na-fluorescein is increased, the physiological pericapillary halo partially destroyed and the interstitial distribution of the dye asymmetrical. In primary lymphedema the lymphatic network filled from the deposit of fluorescent dextran extends much more than in healthy controls. Pathological lymphatic microvessels occur.

Adult↗
PubMed2003

Phosphoinositide-3-kinase-independent contractile activities associated with Fcgamma-receptor-mediated phagocytosis and macropinocytosis in macrophages.

Previous studies have shown that Fcgamma receptor (FcR)-mediated phagocytosis and macropinocytosis in macrophages consist of two dissociable activities: a phosphoinositide 3-kinase (PI3K)-independent extension of phagocytic cups and a PI3K-dependent contractile mechanism that closes phagosomes and ruffles into intracellular organelles. Here, we identify an additional contractile activity that persists in the presence of the PI3K inhibitor wortmannin. ML-7, an inhibitor of myosin-light-chain kinase (MLCK), inhibited FcR-mediated phagocytosis, macropinocytosis and cell movements associated with ruffling. Scanning electron microscopy demonstrated a striking difference in morphology between phagocytic cups in the different inhibitors: whereas phagocytic cups of control cells and wortmannin-treated cells conformed closely to particles and appeared to have constricted them, the phagocytic cups in cells treated with ML-7 were more open. Video microscopy of macrophages expressing green-fluorescent-protein (GFP)-actin fusions revealed that bound IgG-opsonized erythrocytes were squeezed during phagosome formation and closure. In ML-7, GFP-actin-rich protrusions extended outward but failed to squeeze particles. Moreover, in contrast to the effects of PI3K inhibitors, ML-7 markedly reduced ruffle movement, and perturbed circular ruffle formation. These PI3K-independent myosin-II-based contractile activities that squeeze phagocytic cups and curve ruffles therefore represent a third component activity of the actin cytoskeleton during phagocytosis and macropinocytosis.

Actin Cytoskeleton↗