Detection of Apoptotic Cells in Planarians by Whole-Mount TUNEL

3-D organization of fibrinogen receptors using computer reconstruction and whole-mount microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102237 ◽

1988 ◽

Vol 46 ◽

pp. 30-31

Author(s):

E. T. O'Toole ◽

R. R. Hantgan ◽

J. C. Lewis

Keyword(s):

Whole Blood ◽

Colloidal Gold ◽

Blood Platelets ◽

Cell Contact ◽

Computer Assisted ◽

Adherent Cells ◽

Differential Centrifugation ◽

Computer Reconstruction ◽

Sds Page ◽

Whole Mount

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.

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Applications of Scanning Microscopy in Biomedical Research

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100101517 ◽

1968 ◽

Vol 26 ◽

pp. 354-355

Author(s):

T. L. Hayes

Keyword(s):

Information Content ◽

Biomedical Research ◽

Biomedical Applications ◽

Three Dimensional ◽

Dental Enamel ◽

Resolving Power ◽

Whole Mount ◽

Two Parameters ◽

Content Information ◽

Sectioned Tissue

Biomedical applications of the scanning electron microscope (SEM) have increased in number quite rapidly over the last several years. Studies have been made of cells, whole mount tissue, sectioned tissue, particles, human chromosomes, microorganisms, dental enamel and skeletal material. Many of the advantages of using this instrument for such investigations come from its ability to produce images that are high in information content. Information about the chemical make-up of the specimen, its electrical properties and its three dimensional architecture all may be represented in such images. Since the biological system is distinctive in its chemistry and often spatially scaled to the resolving power of the SEM, these images are particularly useful in biomedical research.In any form of microscopy there are two parameters that together determine the usefulness of the image. One parameter is the size of the volume being studied or resolving power of the instrument and the other is the amount of information about this volume that is displayed in the image. Both parameters are important in describing the performance of a microscope. The light microscope image, for example, is rich in information content (chemical, spatial, living specimen, etc.) but is very limited in resolving power.

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The application of video-enhanced constrast/differential interference constrast microscopy in conjunction with whole mount electron microscopy to the study of organelle transport

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100102419 ◽

1988 ◽

Vol 46 ◽

pp. 66-67

Author(s):

J. H. Hayden

Keyword(s):

Electron Microscopy ◽

Rana Pipiens ◽

Silver Film ◽

Immunofluorescence Microscopy ◽

Organelle Transport ◽

Linear Element ◽

Whole Mount ◽

Carbon Coated ◽

Linear Elements ◽

Dic Microscopy

In a previous study, Allen video-enhanced constrast/differential interference constrast (AVEC-DIC) microscopy was used in conjunction with immunofluorescence microscopy to demonstrate that organelles and vesicle move in either direction along linear elements composed of microtubules. However, this study was limited in that the number of microtubules making up a linear element could not be determined. To overcome this limitation, we have used AVEC-DIC microscopy in conjunction with whole mount electron microscopy.Keratocytes from Rana pipiens were grown on glass coverslips as described elsewhere. Gold London Finder grids were Formvar- and carbon coated, and sterilized by exposure to ultraviolet light. It is important to select a Formvar film that gives a grey reflection when it is floated on water. A silver film is too thick and will detract from the image in the light microscope.

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