Use of video cell tracking to identify embryogenic cultured cells

1997 ◽  
pp. 1129-1173 ◽  
Author(s):  
Marcel A. J. Toonen ◽  
Sacco C. de Vries
Keyword(s):  
Cell Tracking ◽  
Cultured Cells

Abstract 1105: Gadofluorine M-Cy 3, a Novel Paramegnetic Contrast Agent for the in vivo and Histolgical Identification of Transplanted Cells.

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Eric Adler ◽  
Anne Bystrup ◽  
Karen Saebo ◽  
Venketesh Mani ◽  
Steve Giovannone ◽  
...  
Keyword(s):  
Iron Oxide ◽  
Fluorescence Microscopy ◽  
Contrast Agent ◽  
Cell Tracking ◽  
Cultured Cells ◽  
Embryonic Stem ◽  
Positive Contrast ◽  
In Vivo Detection ◽  
Gadofluorine M

PURPOSE: Gadofluorine M with a fluorescent dye (GdFMCy3) is a lipophilic paramegnetic contrast agent that is readily absorbed by cultured cells. We hypothesized that this agent would be superior to iron oxide based techniques for cell tracking post cell transplantation. METHODS: Embryonic Stem Cell derived cardiac progenitor cells (ES-CPCs) were generated using previously established methods and incubated for 12 hours with 5 mM GdFMCy3, or transfected with iron oxide using published protocols. Cell survival was >95% for cells incubated with both GdFMCy3 and iron. 500,000 cells labelled with GdFMCy3, iron oxide or control (no contrast agent) were directly injected into the myocardium of mice (n=5/group). Mice were scanned over a two week interval post injection at 9.4T using gated T1-weighted sequences (GdFMCy3), T2* weighted GRE sequences (iron oxide) or the positive contrast sequence GRASP (iron oxide). Mice were sacrificed and the hearts sectioned for microscopy. Perl staining and fluorescence microscopy were used to identify iron oxide and GdFMCy3 within the myocardium, respectively. RESULTS: GdFMCy3 labelled cells were successfully identified in vivo at 9.4T (figure panel B). Good correlation between MRI and histology was observed for both cell labels (figure ). Contrast to noise ratios were significantly higher in the GdFMCy3 group relative to the iron oxide group (figure ). CONCLUSIONS: GdFM-Cy3 is readily taken up by stem cells and easily identified by both MRI and fluorescence microscopy. Given its superior contrast to noise ratio GdFM-Cy3 may be an excellent alternative to iron oxide for in vivo detection and tracking of transplanted cells.

Ultrastructural Correlations between Clonal Human Tumor Colonies and in Vivo Neoplasms

1982 ◽  
Vol 40 ◽  
pp. 210-211
Author(s):  
M.J. Murphy ◽  
R.R. Price ◽  
J.C. Sloman
Keyword(s):  
Cultured Cells ◽  
Human Tumor ◽  
Cell Type ◽  
Tumor Mass ◽  
Initial Cell ◽  
Cloning Assay ◽  
Human Tumor Cloning ◽  
The Relationship

The in vitro human tumor cloning assay originally described by Salmon and Hamburger has been applied recently to the investigation of differential anti-tumor drug sensitivities over a broad range of human neoplasms. A major problem in the acceptance of this technique has been the question of the relationship between the cultured cells and the original patient tumor, i.e., whether the colonies that develop derive from the neoplasm or from some other cell type within the initial cell population. A study of the ultrastructural morphology of the cultured cells vs. patient tumor has therefore been undertaken to resolve this question. Direct correlation was assured by division of a common tumor mass at surgical resection, one biopsy being fixed for TEM studies, the second being rapidly transported to the laboratory for culture.

Electron Probe Microanalysis of Frozen Hydrated Kidneys, Isolated Cells, and Cultured Cells

1982 ◽  
Vol 40 ◽  
pp. 402-403 ◽  
Author(s):  
Claude Lechene
Keyword(s):  
Liquid Nitrogen ◽  
Electron Probe Microanalysis ◽  
Electron Probe ◽  
Cultured Cells ◽  
Liquid Nitrogen Temperature ◽  
Elemental Content ◽  
Isolated Cells ◽  
Renal Tubular Cells ◽  
Diamond Saw ◽  
Saw Blade

Electron probe microanalysis of frozen hydrated kidneysThe goal of the method is to measure on the same preparation the chemical elemental content of the renal luminal tubular fluid and of the surrounding renal tubular cells. The following method has been developed. Rat kidneys are quenched in solid nitrogen. They are trimmed under liquid nitrogen and mounted in a copper holder using a conductive medium. Under liquid nitrogen, a flat surface is exposed by sawing with a diamond saw blade at constant speed and constant pressure using a custom-built cryosaw. Transfer into the electron probe column (Cameca, MBX) is made using a simple transfer device maintaining the sample under liquid nitrogen in an interlock chamber mounted on the electron probe column. After the liquid nitrogen is evaporated by creating a vacuum, the sample is pushed into the special stage of the instrument. The sample is maintained at close to liquid nitrogen temperature by circulation of liquid nitrogen in the special stage.

The Infection of Cells by Bullet-Shaped Viruses

1969 ◽  
Vol 27 ◽  
pp. 372-373
Author(s):  
Frederick A. Murphy ◽  
Alyne K. Harrison ◽  
Sylvia G. Whitfield
Keyword(s):  
Electron Microscopy ◽  
Physical Properties ◽  
Host Cell ◽  
Thin Section ◽  
Cultured Cells ◽  
Cell Infection ◽  
Thin Section Electron Microscopy ◽  
Section Electron ◽  
Section Electron Microscopy

The bullet-shaped viruses are currently classified together on the basis of similarities in virion morphology and physical properties. Biologically and ecologically the member viruses are extremely diverse. In searching for further bases for making comparisons of these agents, the nature of host cell infection, both in vivo and in cultured cells, has been explored by thin-section electron microscopy.

Ultrastructure of two neoplasms in culture compared with original biopsies

1984 ◽  
Vol 42 ◽  
pp. 50-51
Author(s):  
Joseph M. Harb ◽  
James T. Casper ◽  
Vlcki Piaskowski
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Biopsy Specimen ◽  
Cultured Cells ◽  
Light And Electron Microscopy ◽  
Human Tumor ◽  
Soft Agar ◽  
Human Tumor Cells ◽  
Morphological Distinction ◽  
Tumor Types

The application of tissue culture and the newer methodologies of direct cloning and colony formation of human tumor cells in soft agar hold promise as valuable modalities for a variety of diagnostic studies, which include morphological distinction between tumor types by electron microscopy (EM). We present here two cases in which cells in culture expressed distinct morphological features not apparent in the original biopsy specimen. Evaluation of the original biopsies by light and electron microscopy indicated both neoplasms to be undifferentiated sarcomas. Colonies of cells propagated in soft agar displayed features of rhabdomyoblasts in one case, and cultured cells of the second biopsy expressed features of Ewing's sarcoma.

Uultrastructure of Microtubules

1972 ◽  
Vol 30 ◽  
pp. 252-253
Author(s):  
Ariaki Nagayama
Keyword(s):  
Fine Structure ◽  
Cultured Cells ◽  
Present Report ◽  
Confluent Monolayer ◽  
Purification Method ◽  
Vinca Rosea ◽  
L Cells ◽  
Antimitotic Activity ◽  
Monolayer Cultures ◽  
Rapid Purification

Vinblastine(Vb) or vincristine, alkaloid derived from Vinca rosea is known for its antimitotic activity by regrouping of microtubules into paracrystalline form within the cells. A rapid purification method of vinblastine-induced microtubular paracrystals(PC) has provided us with a fresh and pure microtubular material demonstrating the presence of a labile ATPase associated with the PC. The present report is concerned with the fine structure of purified microtubules of mammalian cultured cells.Confluent monolayer cultures of L cells were incubated for 20hrs with 10-5 M Vb (donated from Shionogi Seiyaku & Co., Osaka, Japan).

DNA nick end labeling: a reliable marker for apoptosis?

1995 ◽  
Vol 53 ◽  
pp. 962-963
Author(s):  
Anne F. Bushnell ◽  
Sarah Webster ◽  
Lynn S. Perlmutter
Keyword(s):  
Cell Death ◽  
Cultured Cells ◽  
Apoptotic Cell ◽  
Morphological Characteristics ◽  
Detection Methods ◽  
Tissue Preparation ◽  
Preparation Methods ◽  
Dna Laddering ◽  
Disease States ◽  
Reliable Marker

Apoptosis, or programmed cell death, is an important mechanism in development and in diverse disease states. The morphological characteristics of apoptosis were first identified using the electron microscope. Since then, DNA laddering on agarose gels was found to correlate well with apoptotic cell death in cultured cells of dissimilar origins. Recently numerous DNA nick end labeling methods have been developed in an attempt to visualize, at the light microscopic level, the apoptotic cells responsible for DNA laddering.The present studies were designed to compare various tissue processing techniques and staining methods to assess the occurrence of apoptosis in post mortem tissue from Alzheimer's diseased (AD) and control human brains by DNA nick end labeling methods. Three tissue preparation methods and two commercial DNA nick end labeling kits were evaluated: the Apoptag kit from Oncor and the Biotin-21 dUTP 3' end labeling kit from Clontech. The detection methods of the two kits differed in that the Oncor kit used digoxigenin dUTP and anti-digoxigenin-peroxidase and the Clontech used biotinylated dUTP and avidinperoxidase. Both used 3-3' diaminobenzidine (DAB) for final color development.

Intralysosomal Accumulation of Colloidal Gold-Low Density Lipoprotein Conjugates in Chloroquine-Treated Fibroblasts

1985 ◽  
Vol 43 ◽  
pp. 546-547 ◽  
Author(s):  
Dean A. Handley ◽  
Cynthia M. Arbeeny ◽  
Larry D. Witte
Keyword(s):  
Colloidal Gold ◽  
Cultured Cells ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Coated Vesicle ◽  
Low Density ◽  
Cholesterol Esters ◽  
Cultured Fibroblasts ◽  
Surface Receptors ◽  
Ldl Particles

Low density lipoproteins (LDL) are the major cholesterol carrying particles in the blood. Using cultured cells, it has been shown that LDL particles interact with specific surface receptors and are internalized via a coated pit-coated vesicle pathway for lysosomal catabolism. This (Pathway has been visualized using LDL labeled to ferritin or colloidal gold. It is now recognized that certain lysomotropic agents, such as chloroquine, inhibit lysosomal enzymes that degrade protein and cholesterol esters. By interrupting cholesterol ester hydrolysis, chloroquine treatment results in lysosomal accumulation of cholesterol esters from internalized LDL. Using LDL conjugated to colloidal gold, we have examined the ultrastructural effects of chloroquine on lipoprotein uptake by normal cultured fibroblasts.

Growth factors and the primary cilium in BALB/c 3T3 cells

1987 ◽  
Vol 45 ◽  
pp. 830-831
Author(s):  
R. W. Tucker ◽  
N. S. More ◽  
S. Jayaraman
Keyword(s):  
Growth Factors ◽  
Primary Cilium ◽  
Cultured Cells ◽  
Morphological Changes ◽  
Calcium Ionophore ◽  
Growth Stimulation ◽  
Calcium Ionophore A23187 ◽  
Ionophore A23187 ◽  
3T3 Cells ◽  
Microtubule Depolymerization

The mechanisms by which polypeptide growth factors Induce DNA synthesis in cultured cells is not understood, but morphological changes Induced by growth factors have been used as clues to Intracellular messengers responsible for growth stimulation. One such morphological change has been the transient disappearance of the primary cilium, a “9 + 0” cilium formed by the perinuclear centriole in interphase cells. Since calcium ionophore A23187 also produced both mitogenesis and ciliary changes, microtubule depolymerization might explain ciliary disappearance monitored by indirect immunofluorescence with anti-tubulin antibody. However, complete resorption and subsequent reformation of the primary cilium occurs at mitosis, and might also account for ciliary disappearance induced by growth factors. To settle this issue, we investigated the ultrastructure of the primary cilium using serial thin-section electron microscopy of quiescent BALB/c 3T3 cells before and after stimulation with serum.

A comparison of three cryotechniques (freeze-drying, cryosubstitution, cryosectioning) of specimen preparation for immunolabelling of bacterial antigens of Coxiella burnetii

1994 ◽  
Vol 52 ◽  
pp. 140-141
Author(s):  
T. F. McCaul ◽  
R. J. Gould
Keyword(s):  
Coxiella Burnetii ◽  
Freeze Drying ◽  
Stress Proteins ◽  
Cultured Cells ◽  
High Specificity ◽  
Specimen Preparation ◽  
Bacterial Antigens ◽  
Structural Preservation ◽  
Nuclear Ribonucleoprotein ◽  
Cellular Components

Immuno-electron microscopy has allowed the selective localisation of molecules with high resolution and high specificity. Cryopreparatory methods have provided better retention of antigenicity suitable for precise immunolabelling together with optimal structural preservation of cellular components. Cryosubstitution and cryoultramicrotomy have widely been exploited for immunolabelling. Molecular Distillation Dryer (MDD), a form of freeze-drying technique, has recently been used for immunolabelling of Plasmodium falciparum stress proteins and nuclear ribonucleoprotein particles in cultured cells. In the present study, we report the comparison of all three cryotechniques in the immunolabelling of bacterial antigens of Coxiella burnetii.The highly infectious C. burnetii was prefixed in 3% glutaraldehyde (66 mM cacodylate buffer, pH 6.8 ). The cells were then pre-embedded in 2% low-temperature agarose on Durapore hydrophilic membrane prior to cryofixation using a LifeCell CF100 metal-mirror system. A 1% glutaraldehyde in 100% methanol was used as a medium for cryosubstitution in a Reichert CS Auto Cryosubstitution apparatus.

Sign in / Sign up

Export Citation Format

Share Document