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 685 records · Page 38Linked to original sources
PubMed2002

Automated tri-image analysis of stored corneal endothelium.

BACKGROUND: Endothelial examination of organ culture stored corneas is usually done manually and on several mosaic zones. Some banks use an image analyser that takes account of only one zone. This method is restricted by image quality, and may be inaccurate if endothelial cell density (ECD) within the mosaic is not homogeneous. The authors have developed an analyser that has tools for automatic error detection and correction, and can measure ECD and perform morphometry on multiple zones of three images of the endothelial mosaic. METHODS: 60 human corneas were divided into two equal groups: group 1 with homogeneous mosaics, group 2 with heterogeneous ones. Three standard microscopy video images of the endothelium, graded by quality, were analysed either in isolation (so called mono-image analysis) or simultaneously (so called tri-image analysis), with 50 or 300 endothelial cells (ECs) counted. The automated analysis was compared with the manual analysis, which concerned 10 non-adjacent zones and about 300 cells. For each analysis method, failures and durations were studied according to image quality. RESULTS: All corneas were able to undergo analysis, in about 2 or 7.5 minutes for 50 and 300 ECs respectively. The tri-image analysis did not increase analysis time and never failed, even with mediocre images. The tri-image analysis of 300 ECs was always most highly correlated with the manual count, particularly in the heterogeneous cornea group (r=0.94, p<0.001) and prevented serious count errors. CONCLUSIONS: This analyser allows reliable and rapid analysis of ECD, even for heterogeneous endothelia mosaics and mediocre images.

Aged↗
PubMed2003

Dynamic assessment of angiogenesis in renal cell carcinoma spheroids by intravital microscopy.

PURPOSE: Renal cell cancer represents a suitable tumor model for in vivo observation of neo-angiogenesis. We used intravital microscopy and the well established dorsal skin fold chamber model to characterize neo-angiogenesis in freely implanted renal cell cancer spheroids. MATERIAL AND METHODS: Tumor spheroids were implanted into dorsal skin fold chambers of 8 nude mice. At days 3, 6, 10 and 14 after implantation the newly vascularized spheroid area, density of perfused microvessels in the spheroid versus the periphery, capillary center erythrocyte velocity and capillary diameter were recorded by intravital microscopy. Video images were analyzed by a computer assisted image analysis device. After the experiments the chambers were analyzed morphologically. RESULTS: The model enabled quantitative analysis of microcirculation and angiogenesis in the renal cell cancer spheroids during 14 days of observation. Mean spheroid center perfused microvessel density +/- SEM increased from 3 +/- 2 to 269 +/- 21 cm.-1 on days 3 to 10 and subsequently decreased to 189 +/- 38 cm.-1 on day 14. Spheroid periphery perfused microvessel density was significantly higher throughout the experiments, attaining a mean maximum of 522 +/- 34 cm. on day 14. Mean capillary diameter decreased continuously from 14.2 +/- 0.9 to 8.4 +/- 0.4 microm. on days 3 to 14. In contrast, mean capillary center erythrocyte velocity significantly increased during 14 days of observation from 0.09 + 0.02 mm. per second on day 3 to 0.24 +/- 0.08 mm. per second on day 14. Histological analysis after 14 days revealed the spheroids as cell clusters in the upper layers of the dorsal skin fold chamber. CONCLUSIONS: The model is suitable for the analysis of renal cell cancer angiogenesis. Although it is heterotopic, angiogenesis in renal cell cancer spheroids mimics important characteristics of human renal cell cancer.

Animals↗
PubMed2000

Detection of leukocytes in contact with the vessel wall from in vivo microscope recordings using a neural network.

Leukocytes play an important role in the host defense as they may travel from the blood stream into the tissue in reacting to inflammatory stimuli. The leukocyte-vessel wall interactions are studied in post capillary vessels by intravital video microscopy during in vivo animal experiments. Sequences of video images are obtained and digitized with a frame grabber. A method for automatic detection and characterization of leukocytes in the video images is developed. Individual leukocytes are detected using a neural network that is trained with synthetic leukocyte images generated using a novel stochastic model. This model makes it feasible to generate images of leukocytes with different shapes and sizes under various lighting conditions. Experiments indicate that neural networks trained with the synthetic leukocyte images perform better than networks trained with images of manually detected leukocytes. The best performing neural network trained with synthetic leukocyte images resulted in an 18% larger area under the ROC curve than the best performing neural network trained with manually detected leukocytes.

Animals↗
PubMed1993

Gastrulation and mesoderm morphogenesis in the white sturgeon.

This study presents a detailed description of gastrulation in the white sturgeon, Acipenser transmontanus, using scanning electron microscopy, histology, and time-lapse filming and video microscopy. This morphological analysis describes the similarity of gastrula structure in the sturgeon and the amphibian Xenopus laevis, and suggests that the species share many developmental mechanisms. It also identifies important differences, such as the equatorial dorsal lip in sturgeon, and provides a basis for interpreting experiments that test the effect of these differences on gastrulation. The onset of gastrulation in the sturgeon is marked by the appearance of a blastoporal equatorial pigment line that forms as the apices of bottle cells contract and concentrate surface pigment granules. Bottle cell formation at the blastopore lip and involution of surface material through the blastopore are strikingly similar to the equivalent processes in amphibian embryos. As gastrulation continues, a distinct cleft of Brachet forms between pre-involution and post-involution material. Following involution, the prospective axial mesoderm located on the dorsal surface of the late blastula (Ballard and Ginsburg: J. Exp. Zool., 213:69-103, 1980) ingresses from a central zone in the posterior archenteron roof surface in a process that is unlike any in Xenopus, but resembles events in other amphibians (Purcell, 1992; Smith: Dev. Biol., 98:250-254, 1983; King: Biol. Bull., 4:287-300, 1903). The detailed comparison of similarities and differences in gastrulation in different vertebrate lineages yields insights into the function and versatility of common developmental mechanisms.

Animals↗
PubMed1992

Minipodia, novel structures for extension of the lamella: a high-spatial-resolution video microscopic study.

Extension of leading lamellae has been analyzed by high-spatial- and -time-resolution video microscopy. Many tiny semicircular profiles, hereby named minipodia, have been found at the leading edge of locomoting fibroblasts in culture. These new structures are not "miniruffles" since minipodia seem to originate underneath the leading lamella. Once initiated, minipodia quickly expand and become indistinguishable from what have been known as lamellipodia. The minipodial front becomes the new focal leading edge. So, it appears that leading lamellae do not extend directly themselves, but minipodia protrude underneath them to become new leading edges. These findings are not compatible with the current concept of cell movement, and a new model is proposed.

3T3 Cells↗
PubMed1992

Colour thresholding and objective quantification in bioimaging.

Computer imaging is rapidly becoming an indispensable tool for the quantification of variables in research and medicine. Whilst its use in medicine has largely been limited to qualitative observations, imaging in applied basic sciences, medical research and biotechnology demands objective quantification of the variables in question. In black and white densitometry (0-256 levels of intensity) the separation of subtle differences between closely related hues from stains is sometimes very difficult. True-colour and real-time video microscopy analysis offer choices not previously available with monochrome systems. In this paper we demonstrate the usefulness of colour thresholding, which has so far proven indispensable for proper objective quantification of the products of histochemical reactions and/or subtle differences in tissue and cells. In addition, we provide interested, but untrained readers with basic information that may assist decisions regarding the most suitable set-up for a project under consideration. Data from projects in progress at Tulane are shown to illustrate the advantage of colour thresholding over monochrome densitometry and for objective quantification of subtle colour differences between experimental and control samples.

Animals↗
PubMed1980

Microvessel mean transit time and blood flow velocity of sulfhemoglobin-RBC.

An indicator dilution technique is described for obtaining time-concentration curves subsequent to bolus injections of sulfhemoglobin red blood cells (SH-RBC), which have a deep greenish-brown color (absorption peak 620 nm vs. 542 and 564 nm for normal red cells). The series- and parallel-coupled microvessels of cat mesentery were studied. This is accomplished by means of video microscopy with a two-window intensity-sensitive video sampler system. The relationship between SH-RBC concentration in blood and optical measurement is linear. Blood flow velocities were calculated from the difference in mean transit times between two points along a vessel. When this technique is used in association with the previously reported method for determining time-concentration curves for the plasma indicator FITC-dextran the mean transit time (t) for red blood cells was less than for plasma in arterioles. The reproducibility of t and flow velocity for both SH-RBC and FITC-dextran from successive injections were reported. The mean transit time ratio of arteriolar SH-RBC to FITC-dextran averages 0.89. Blood flow velocity calculated from SH-RBC is greater than that calculated from FITC-dextran in these same arterioles. The ratio of the velocities averages 1.29.

Animals↗
PubMed2004

Basal sliding and the mechanics of oscillation in a mammalian sperm flagellum.

The mechanism of oscillation in cilia and flagella has been a long-standing mystery. This article raises the possibility of a mechanical explanation based on new findings relating to where in the flagellum microtubule sliding can occur--and where it cannot occur. All theoretical analyses of flagellar bending have until now made the assumption that sliding displacements at the base of the flagellum cannot occur. One consequence of this has been the need to accept that sliding must be transmitted through propagating bends, an idea that has been tolerated even though it becomes paradoxical if bends are the result of resistance to sliding. Our observations, of spermatozoa from the chinchilla, have led us to a contradictory view. We have shown directly, by light microscopy and by two methods of electron microscopy, that basal sliding does occur. Also, evidence from video microscopy indicates that a propagating bend cannot transmit sliding through it. We have analyzed a movement pattern in which the beat frequency increases fourfold in a phasic manner. Our analysis of this suggests that new bends terminate when no further sliding is possible. At this point the bend direction immediately reverses. That is, the flagellar beat frequency increases when there is a limitation to sliding. One can see directly the alternation in basal sliding direction under these circumstances. This suggests a mechanism for the initiation of a new bend in the opposite direction to the bend just completed: we propose that the initiating trigger is the reversal of elastic deformations at the base, which reverses the direction of interdoublet sliding.

Animals↗
PubMed1993

Multivesicular bodies in HEp-2 cells are maturing endosomes.

Conventional fluorescence microscopy of fixed HEp-2 cells as well as video microscopy of living cells incubated with transferrin-Texas Red (Tf-TxR) for < 60 min revealed distinct punctuate endosomal structures. Quantitative ultrastructural analysis using horseradish peroxidase (HRP) and cationized gold as tracers showed that spherical multivesicular bodies (MVBs) were the predominant endocytic compartments in HEp-2 cells and that MVBs within 60 to 90 min matured into lysosomes still containing internal vesicles. The number of labeled MVBs increased continuously from 2.5 min to 30 min of tracer incubation. However, when the cells were pulsed for 5 min followed by 10 or 25 min chases, the number of labeled MVBs corresponded to that obtained after 5 min of continuous incubation. The diameter of labeled MVBs was largely constant with time, but the number of internal MVB vesicles increased. Thus, early or newly formed MVBs contained few internal vesicles, whereas late MVBs, that is to say, MVBs that have existed for some period of time, contained numerous internal vesicles, and finally a mixture of membranous material or myelin figures and vesicles. It is thus in principle possible to distinguish between early and late MVBs in HEp-2 cells on the basis of morphology. However, the difference in number of internal vesicles applies only to the entire MVB population; after only 2.5 to 5 min of incubation, MVBs with numerous internal vesicles could also be reached by internalized tracer. Concomitant with the gradual changes in morphology, the MVBs also showed a characteristic change in content of marker proteins as detected by immunogold labeling on ultracryosections. Hence, early MVBs with relatively few internal vesicles and typically reached by internalized tracers within 5 min contained transferrin receptors (TfRs). By contrast, MVBs with many internal vesicles and labeled after 60 min of incubation contained mannose-phosphate receptors (MPRs), and the MVBs with distinct membranous material or myelin figures in addition to the internal vesicles were enriched in the lysosome membrane protein lamp-1. Thus, there seems to be a gradual maturation of MVBs in HEp-2 cells.

Carcinoma, Squamous Cell↗
PubMed2002

The role of histamine in dural vessel dilation.

The pain of migraine is often throbbing suggesting an important role for the cranial blood vessels and their innervation by the trigeminal nerve. It is proposed that clinically effective anti-migraine compounds, such as 5-HT(1B/1D) agonists, have actions that include inhibiting calcitonin gene-related peptide (CGRP) release from trigeminal nerves. Human studies suggest that histamine can induce migraine possibly by activating nitric oxide (NO) synthase to promote endogenous NO production. The present studies investigated the effect of histamine and its antagonists on the cranial blood vessels using intravital microscopy to assess directly the diameter of dural arteries in sodium pentobarbitone anaesthetised rats. Electrical stimulation of a closed cranial window produces, by local depolarisation of nerves, dural vessel dilation that is monitored continuously on-line using video-microscopy and a video dimension analyser. Histamine infusion caused immediate and reproducible dilation of meningeal blood vessels (103.5+/-6%; n=40) that could be blocked by H(1)- (mepyramine) and H(2) (famotidine)-receptor antagonists (P<0.05), as well as a nitric oxide synthase inhibitor (N(G)-nitro-L-arginine methylester; P<0.05). Neurogenic dural vasodilation was not inhibited by H(2)-receptor antagonists, but was significantly inhibited by a H(1)-receptor antagonist at the high dose of 10 mg/kg. The present studies demonstrate that histamine is likely to activate NO synthase to promote NO production. There is also evidence that H(1)-receptors may be present on trigeminal neurones as the H(1)-receptor antagonist inhibited neurogenic vasodilation, albeit at a large dose.

Animals↗
PubMed2001

The effect of adrenergic compounds on neurogenic dural vasodilatation.

The pharmacology of neurogenic trigeminovascular vasodilator responses in the dura mater is of interest for understanding the pathophysiology of migraine and to develop new therapies for this disabling common condition. Aminergic mechanisms have been implicated in migraine through direct study of amines in patients, and by inference from the pharmacology of many effective anti-migraine compounds, particularly preventative agents. This study used intravital microscopy to assess the role of aminergic transmission in neurogenic dural vasodilatation (NDV) by measuring directly the diameter of dural arteries in sodium pentobarbitone anaesthetised rats. Electrical stimulation of a closed cranial window produces, by local depolarisation of nerves, dural vessel dilation that is monitored continuously on-line using video-microscopy and a video dimension analyser. This dural vasodilatation was not affected by pre-treatment with an alpha1-adrenoceptor agonist (phenylephrine, 1 and 5 microg/kg), or antagonist (corynanthine, 1 and 2 mg/kg), nor by an alpha2-adrenoceptor agonist (UK14,304, 5 microg/kg) or antagonist (yohimbine, 1 and 3 mg/kg). Similarly, we saw no effect of beta-adrenoceptor blockade (propranolol, 1 and 3 mg/kg). The lack of an inhibitory effect of UK14,304 the model of neurogenic dural vasodilation contrasts with its effect in neurogenic dural plasma protein extravasation model. The lack of inhibition of beta-adrenoceptor antagonists in the neurogenic vasodilatation model contrasts with their usefulness as migraine prophylactics, and suggests that their mechanism of action in migraine is unlikely to be through sensory trigeminal fibre terminals at the neurovascular junction. Moreover, the data indicate that the adrenergic system does not play a significant role in neurogenic dural vasodilation.

Adrenergic Agents↗
PubMed1990

A delay in membrane fusion: lag times observed by fluorescence microscopy of individual fusion events induced by an electric field pulse.

Low light level video microscopy of the fusion of DiI- (1,1'-dihexadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate) labeled rabbit erythrocyte ghosts with unlabeled rabbit erythrocyte ghosts, held in stable apposition by dielectrophoresis in sodium phosphate buffers, showed reproducible time intervals (delays) between the application of a single fusogenic electric pulse and the earliest detection of fluorescence in the unlabeled adjacent membranes. The delay increased over the range 0.3-4 s with a decrease in (i) the electric field strength of the fusion-inducing pulse from 1000 to 250 V/mm, (ii) the decay half-time of the fusogenic pulse in the range 1.8-0.073 ms, and (iii) the dielectrophoretic force which brings the membranes into close apposition. A change in the buffer viscosity from 1.8 to 10 mP.s caused the delay to increase from 0.36 to 3.7 s (in glycerol solutions) or to 5.2 s (in sucrose solutions). The delay decreased 2-3 times with an increase in temperature from 21 to 37 degrees C. It did not differ significantly for "white" ghosts [0.013 mM hemoglobin (Hb)] or "red" ghosts (0.15 mM Hb) or buffer strength over the range 5-60 mM (sodium phosphate, pH 8.5). The calculated activation energy, 17 kcal/mol, does not depend on the field strength. The yield of fused cells was high when the delay was short. The delay in electrofusion resembles the delays in pH-dependent fusion of vesicular stomatitis viruses with erythrocyte ghosts [Clague, M. J., Schoch, C., Zech, L., & Blumenthal, R. (1990) Biochemistry 29, 1303-1308] and of fibroblasts expressing influenza hemagglutinin and red blood cells [Morris, S. J., Sarkar, D.P., White, J. M., & Blumenthal, R. (1989) J. Biol. Chem. 264, 3972-3978].(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Functional sorting of actin isoforms in microvascular pericytes.

We characterized the form and distribution of muscle and nonmuscle actin within retinal pericytes. Antibodies with demonstrable specificities for the actin isoforms were used in localization and immunoprecipitation experiments to identify those cellular domains that were enriched or deficient in one or several actin isoforms. Living pericyte behavior was monitored with phase-contract video microscopy before fixation to identify those cellular areas that might preferentially be stained with either of the fluorescent antiactins or phallotoxins. Antibody and phallotoxin staining of pericytes revealed that nonmuscle actin is present within membrane ruffles, pseudopods, and stress fibers. In contrast, muscle actin could be convincingly localized in stress fibers, but not within specific motile areas of pericyte cytoplasm. To confirm and quantitatively extend the results obtained by fluorescence microscopy, nonionic and ionic detergents were used to selectively extract the motile or immobilized (stress fiber-containing) regions of biosynthetically labeled pericyte cytoplasm. Immunoprecipitated actins that were present within these discrete cellular domains were subjected to isoelectric focusing in urea-polyacrylamide gels before fluorographic analysis. Scanning laser densitometry of the focused actins could not reveal any detectable alpha-actin within those beta- and gamma-actin-enriched motile regions extracted with nonionic detergents. Moreover, when pericyte stress fibers are completely dissolved by ionic detergent lysis, three actin isoforms can be quantified to be present in a ratio of 1:2.75:3 (alpha:beta:gamma). These biochemical findings on biosynthetically labeled and immunoprecipitated pericyte actins confirm the fluorescent localization studies. While the regulatory events governing this actin sorting are unknown, it seems possible that such events may play important roles in controlling cell shape, adhesion, or the promotion of localized cell spreading.

Actin Cytoskeleton↗
PubMedDate not supplied

Morphologic differences in leukemic cells (L1210) exposed to 1-beta-D-arabinofuranosylcytosine and daunorubicin.

To compare the morphologic differences in leukemic cells exposed to a nucleotide synthesis inhibitor and a DNA strand damage inducer, L1210 cells incubated with 1-beta-D-arabinofuranosylcytosine (ara-C) or daunorubicin (DNR) were investigated by time-lapse video microscopy, flow cytometry and transmission electron microscopy. At 1.0 microgram/ml of ara-C, cells could be separated into two populations, with some becoming ballooned and deformed, i.e. irreversibly damaged, and others remaining unchanged, i.e. in a proliferative state. In contrast, at 0.10 microgram/ml of DNR, nearly all cells were remarkably enlarged and did not proliferate during 72 hours of observation. Such morphologic changes induced at clinically achievable concentrations of each agent seemed to reflect their differing mechanisms of action on the synthesis of DNA. The methods described may allow the characterization of antileukemic agents as well as the development of clinical designs for their rational application in leukemia patients.

Animals↗
PubMed1993

Organelle movements in the wild type and wall-less fz;sg;os-1 mutants of Neurospora crassa are mediated by cytoplasmic microtubules.

The cellular basis of organelle transport in filamentous fungi is still unresolved. Here we have studied the intracellular movement of mitochondria and other organelles in the fungus Neurospora crassa. Four different model systems were employed: hyphae, protoplasts, a cell wallless mutant, and experimentally generated small, flattened cell fragments of the mutant cells. Organelle movements were visualized by DIC optics and computer-enhanced video microscopy. In all cell models the transport of organelles was vectorial and saltatory in nature. The mean velocities for mitochondria, particles and nuclei were 1.4, 2.0, and 0.9 microns/s, respectively. Treatment with 10 microM nocodazole for 30 minutes caused a complete disappearance of microtubules and reversibly blocked directed transport of virtually all organelles, whereas cytochalasin D up to 20 microM was without effect. Correlative video and immunofluorescence microscopy of small fragments of wall-less mutant cells revealed a clear match between microtubule distribution and the tracks of moving organelles. We conclude that organelle movement in the filamentous fungus Neurospora crassa is a microtubule-dependent process.

Actins↗
PubMed1991

Dual-view microscopy with a single camera: real-time imaging of molecular orientations and calcium.

A new microscope technique, termed "W" (double view video) microscopy, enables simultaneous observation of two different images of an object through a single video camera or by eye. The image pair may, for example, be transmission and fluorescence, fluorescence at different wavelengths, or mutually perpendicular components of polarized fluorescence. Any video microscope can be converted into a dual imager by simple insertion of a small optical device. The continuous appearance of the dual image assures the best time resolution in existing and future video microscopes. As an application, orientations of actin protomers in individual, moving actin filaments have been imaged at the video rate. Asymmetric calcium influxes into a cell exposed to an intense electric pulse have also been visualized.

Actin Cytoskeleton↗
PubMed1988

Structural analysis of the mitotic cycle in pre-gastrula Xenopus embryos.

The long-known phenomenon of karyomere (chromosome vesicle) formation at early telophase of the nuclear cycle during early embryogenesis of a wide range of organisms including amphibians (Rubaschkin 1905; for review, see Richards 1917) was investigated in the early cleavage cycles of Xenopus laevis embryos before the mid blastula transition. Embryos were fixed and Epon embedded at successive time intervals and consecutive thick (3 micron) and ultrathin sections cut. Using conventional light microscopy at low magnification as well as phase and/or interference contrast video microscopy at high magnification, a substantial amount of information could be obtained from the analysis of optical sections in thick-sectioned material. In addition, details of the ultrastructural organization could be analysed from corresponding ultrathin sections by electron microscopy. The light microscopic analysis of serial thick sections allowed precise determination of the arrangement and sizes of telophase karyomere structures during the embryonic nuclear division cycle. It was found that small, widely spaced 1st order karyomeres fuse to larger (2nd order) karyomeres which then progressively exhibit lateral fusion of neighbouring karyomeres. The final coalescence of adjacent karyomeres marks the onset of the reorganization of the typical interphase nuclear structure. The data are discussed with regard to the occurrence of karyomeres during the embryonic nuclear cycle of arthropods, dipteran insects, and echinoderms as well as recent progress in the use of Xenopus egg extracts for in vitro assembly of nuclear structures around protein-free DNA.

Animals↗
PubMed1984

The nasal mucociliary apparatus. Correlation of structure and function in the rat.

Rats are used extensively in inhalation toxicologic studies, but little information is available on the rat nasal mucociliary apparatus. The function of the rat nasal mucociliary apparatus was studied in vitro by video analysis, and its structure was examined by light and electron microscopy. Video analysis permitted determination of mucus flow rate and ciliary beat frequency without disrupting mucociliary function. In different regions of the nose, there was a characteristic mucus flow rate, flow pattern, and ciliary beat frequency, with minimal interanimal variation. Analysis of data pairs revealed no evidence of a correlation between mucus flow rate and ciliary beat frequency, ciliary length, density of the ciliated cell population, or the products of these factors. Mucus was seen to flow over certain nonciliated areas, and it is proposed that the mucus is pulled over these areas. Mucus flow characteristics indicated that the hypophase was less viscous than, and moved in the same direction as, the epiphase. The nasal mucus was present as a continuous layer over the respiratory epithelium, and had an osmiophilic surface film, which was found to be made up of 2 separate membranous components. On the basis of these studies, it was concluded that the rat nasal mucociliary apparatus represents a useful and readily accessible model for in vitro studies of nasal mucociliary function.

Animals↗